JP2017078030A - Cognitive function-improving agent comprising docosahexaenoic acid - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cognitive function-improving agent having the effects of solving the problem of an effective amount of DHA intake relevant to the upper limit of intake and the problem of an excessive amount of intake, exerting the functionality of DHA to brain functions, etc., functioning effectively at safe intake levels, being easily ingested for a long time by reducing intake per day.SOLUTION: The present invention provides a cognitive function-improving agent that comprises docosahexaenoic acid and astaxanthin. The cognitive function-improving agent is used so that docosahexaenoic acid in dose of 200 mg or more and 600 mg or less per day is orally administered to a human.SELECTED DRAWING: Figure 7

Description

  The present invention relates to a cognitive function improving agent containing docosahexaneenoic acid (hereinafter sometimes abbreviated as “DHA”).

  DHA is an n-3 linear polyunsaturated fatty acid having 22 carbon atoms and 6 unsaturated bonds, and is an essential fatty acid contained in fish and the like. In vivo, it is abundant in brain, nerve tissue and sperm. Eicosapentaenoic acid (EPA) contained in fish oil widely used as a DHA source is an n-3 linear polyunsaturated fatty acid having 20 carbon atoms and 5 unsaturated bonds.

  DHA is expected to prevent or improve arteriosclerosis, dyslipidemia, dementia, atopy, allergy, etc. It is also said to suppress cancer development and metastasis. In pregnancy and breastfeeding, it is desirable to take an appropriate amount of DHA for the proper growth of the fetus and infant. As a food, a special health food with DHA and EPA as the ingredients concerned was approved in 2003 with the indication "suitable for those who are concerned about neutral fat", and now it is used for soft drinks and sausages. It is sold in the form (Non-Patent Documents 1 and 2).

  The memory learning function is one of the physiological activities of DHA, and when DHA was administered to a model rat of Alzheimer's disease, a report that the group administered DHA showed improved learning ability compared to the control group in animal studies, It has been reported that reduced learning ability improves. In humans, MMSE (Mini Mental State Examination) is reported to be able to significantly suppress the decline of cognitive function when administered for 6 months with DHA and EPA supplements to patients with very mild Alzheimer's disease with 27 or more points Has been. From these facts, DHA is expected to have an effect on the brain (Non-Patent Documents 3 to 5).

  Foods have nutrition as a primary function and taste as a secondary function. In recent years, the effects on health have been attracting attention as a tertiary function in addition to this. Functional foods containing DHA as an active ingredient have also been studied for the purpose of this tertiary function (Patent Documents 1 and 2). For example, Patent Document 1 relates to a tablet containing various components having antioxidant ability such as DHA, EPA, astaxanthin, lecithin, vitamin E and the like. Patent Document 2 relates to the use of krill oil for pharmaceuticals for lowering cholesterol and LDL in patients, and describes that KHA oil contains DHA and the like. In addition, the addition of antioxidants is also described.

  While DHA and EPA are expected to have various functions, the possibility of health damage due to excessive intake has been pointed out (Non-Patent Document 6). As a specific intake, it has been pointed out that ingestion of 3 g or more per day for an adult may cause a health hazard. In addition, in the US FDA's limited health labeling standard, intake from so-called supplements such as capsule tablets and the like, combined with DHA and EPA, should not exceed 2 g per day for adults. In addition, supplements containing DHA and EPA are mainly produced using fish oil, but as side effects of overdose, belching, nausea, nosebleed, and loose stool have been reported.

  The Ministry of Health, Labor and Welfare recommends ingesting a total of 1 g or more of DHA and EPA per day. However, according to the data compiled by the Ministry of Health, Labor and Welfare, the 50th percentile value of national DHA and EPA intake is 0.260 g per day, The total of 0.128 g is estimated to be 0.388 g. This value is calculated based on the lipid intake in the National Health and Nutrition Survey in 2005 and 2005. The lipid intake in 2005 is from 12.5 g (percentile value) to 134. 3g (99th percentile value) covers a wide range, and it is considered that the DHA and EPA intakes of the people vary greatly between individuals.

  On the other hand, beneficial effects on brain function have been suggested when the intake amount exceeds about 1 g per day as a specific effective amount of DHA or EPA for humans. For example, in a double-blind, randomized, placebo-controlled study of 176 healthy young adults with low DHA intake (18-45 years old, study group 85, New Zealand), DHA 1.16 g / day for 6 months There is a report that improvement was observed only in 2 out of 7 cognitive function tests when ingested for a while (Non-patent Document 7).

Utility Model Registration No. 3192948 Patent No. 5228185

Health food ingredient scientific demonstration database HP: https://www.hfs-data.jp/ National Institute of Health and Nutrition “Health Food” Safety and Efficacy Information HP: http://hfnet.nih.go.jp/ Kazuyoshi Yazawa, Haruo Kageyama, Review Bioactivity of Docosahexaenoic Acid, Oil Chemistry, 1991, 40, 10, 974-978 Hashimoto M., et al. Chronic administration of docosahexaenoic acid ameliorates the impairment of spatial cognition learning ability in amyloid beta-infused rats.J. Nutr. 2005; 135 (3): 549-55. Freund-Levi Y., et al. Omega-3 fatty acid treatment in 174 patients with mild to moderate Alzheimer disease: OmegAD study: a randomized double-blind trial. Arch. Neurol. 2006; 63 (10): 1402-8. Health Food Database First Publication Pharmacist's Letter / Prescriber's Letter Editors (National Institute of Health and Nutrition) Stonehouse W., et al. DHA supplementation improved both memory and reaction time in healthy young adults: a randomized controlled trial. Am J Clin Nutr. 2013 May; 97 (5): 1134-43.

  Here, if it is required to ingest 1 g of DHA, there is a high possibility that EPA is also ingested about 0.5 to 1 g at the same time as the DHA, so the total intake amount does not exceed 2 g or 3 g. It is difficult to control specific intake because of the upper limit of intake that is better. That is, it is considered necessary to control the intake amount as low as possible even within a range of about 1 to 3 g per day, which has been conventionally used as an intake amount of DHA or EPA for obtaining an effective effect on brain function. .

  In addition, the intake amount of 1 to 3 g per day is an amount necessary to ingest 6 to 18 capsules of a commercially available general capsule tablet (DHA content of about 150 mg to 170 mg / capsule) every day. . And the range of such an intake is the range of the intake (preferably 2 g / day or less) which can ensure safety, and the intake (about 1 g / day or more) for obtaining functionality. Nearby and also have some overlapping problems. Moreover, since DHA / EPA contains fish oil, there may be some puffiness, and it is not realistic to take such a large amount of capsules every day. In addition, as mentioned above, it is thought that there is a large individual difference in the amount of daily intake of DHA and EPA of the people, so in addition to the normal diet, food containing 1 g or more of DHA per day Ingestion is expected to have adverse health effects due to chronic overdose.

  On the other hand, Patent Documents 1 and 2 propose that an antioxidant such as carotenoid is arbitrarily selected and combined with an unsaturated fatty acid or the like, but specific combinations of each antioxidant are ingested. The relationship with the quantity has not been fully examined. Under such circumstances, an object of the present invention is to provide a cognitive function improving agent that functions effectively even when DHA is orally administered at a low dose.

  The present invention solves the problem of the effective amount of DHA intake and the upper limit of intake as described above, and the problem of a large amount of intake by the present inventors, etc., and exhibits the functionality of DHA to brain functions and the like. It was discovered as a result of examination.

  From the test described later for finding the present invention, the present inventors orally administrated in the form of a formulation in which DHA and astaxanthin were mixed, and the amount of DHA intake that was not considered effective in the conventional knowledge It has been found that there is an effect of improving cognitive function.

  DHA and EPA are susceptible to oxidation even after being absorbed into the air and body, and DHA and EPA taken orally are considered to be difficult to function efficiently. From the viewpoint of suppressing the influence of this oxidation, it is also conceivable that astaxanthin functions as an antioxidant. However, the effect found in the present invention is based on a characteristic result relating to the effect of improving cognitive function, and can be considered to have an improvement effect on all that is immediately proposed as a function of DHA. However, it cannot be explained from the additional effect of mixing with what can function as an antioxidant. In addition, the effect of the present invention is not appropriately selected as an arbitrary antioxidant, but relates to a specific dose recognized only as a combination with astaxanthin.

That is, the present invention relates to the following inventions.
<1> A cognitive function improving agent containing docosahexaenoic acid (DHA) and astaxanthin, wherein 200 mg or more and 600 mg or less of DHA per day is orally administered to a human. An agent for improving cognitive function, characterized by being used.
<2> The cognitive function improving agent contains eicosapentaenoic acid (EPA), and the weight ratio (DHA: EPA) of DHA and EPA contained in the cognitive function improving agent is 20: 1 to 1. The cognitive function improving agent according to <1>, which is 5: 1.
<3> The cognitive function improving agent according to <1> or <2>, wherein the cognitive function improving agent is a tablet containing 30 mg to 200 mg of DHA per tablet.

  The cognitive function improving agent of the present invention functions effectively with an intake that can be safely ingested, and has an effect of being easily ingested for a long period of time by reducing the intake per day.

It is a front view of the soft capsule prepared as one aspect | mode of the cognitive function improving agent of this invention. It is sectional drawing of the soft capsule prepared as one aspect | mode of the cognitive function improving agent of this invention. It is the figure which put together the test subject's background factor in the Example of this invention. It is a figure which shows transition of the measured value of a brain function (word memory test), and the variation | change_quantity from before ingestion. It is a figure which shows transition of the measured value of a brain function (word recall test), and the variation | change_quantity from before ingestion. It is a figure which shows the variation | change_quantity of a brain function (Stroop test). It is a figure which shows transition of the measured value of a brain function (Japanese version revermede action memory test), and the variation | change_quantity from before ingestion. It is the figure which put together the background factor of the test subject who performed the stratified analysis. It is a figure which shows transition of the measured value of the brain function (Japanese version revermede action memory test | inspection) which performed the stratified analysis, and the variation | change_quantity from before ingestion.

  DESCRIPTION OF EMBODIMENTS Embodiments of the present invention will be described in detail below. However, the description of constituent elements described below is an example (representative example) of an embodiment of the present invention, and the present invention is described below unless the gist thereof is changed. It is not limited to the contents.

  The cognitive function improving agent of the present invention is a cognitive function improving agent containing DHA and astaxanthin, and 200 mg or more and 600 mg or less of DHA per day is orally administered to the human. It is related with the cognitive function improving agent characterized by being used like this. The daily intake of DHA is more preferably 400 mg or more and 600 mg or less. By adopting such a configuration, even if the DHA intake is continued, the cognitive function improving effect can be obtained even if the dose is lowered so that it can be safely ingested. As described above, DHA and EPA are easily affected by oxidation even after being absorbed in the air and in the body, and DHA and EPA taken orally are considered to be difficult to function efficiently and suppress the effects of this oxidation. Astaxanthin may function as an antioxidant from the point of view, but the effect of the present invention is not appropriately selected as an arbitrary antioxidant, but relates to a specific dose recognized only as a combination with astaxanthin. It is.

  Since the DHA raw material generally contains EPA together with DHA, when the amount of DHA administered orally as this cognitive function improving agent exceeds 600 mg, it is common to consider the amount of EPA that is likely to be taken at the same time. The amount taken together with the amount of DHA and EPA (388 mg (50th percentile value)) taken from normal foods in the diet exceeds 1 g, which also starts to cause health hazards. Depending on the intake, a larger amount may be taken chronically, which may result in an excessive intake. The amount of DHA that is orally administered as a cognitive function improving agent is managed as a daily dose for humans, and although it is not necessary to specify a strict relationship with the human body weight, it is basically 60 kg body weight. Therefore, from this amount, the intake per body weight is controlled as 3.3 mg / kg body weight to 10 mg / kg body weight, preferably 6.6 mg / kg body weight to 10 mg / kg body weight. You can also

"Docosahexaenoic acid (DHA)"
DHA is an n-3 linear polyunsaturated fatty acid having 22 carbon atoms and 6 unsaturated bonds, and EPA having 20 carbon atoms and 5 unsaturated bonds. Since DHA is abundant in fish oil, fish oil and some kinds of algae purified are widely used in food. In the present invention, the amount of DHA contained in these fish oil and some kinds of algae is also measured. It can be managed and used in a cognitive function improving agent. These fish oils and some algae generally contain eicosapentaenoic acid (EPA) at the same time, so let's separate DHA and EPA from fish oil and several algae with higher accuracy. Since it may not be realistic, refined fish oil or several kinds of algae containing DHA and EPA may be used.

  The weight ratio (DHA: EPA) of DHA and EPA contained in the cognitive function improving agent of the present invention is preferably 20: 1 to 5: 1, and preferably 20: 1 to 8: 1. More preferred. Since EPA is considered to be unable to break through the blood-brain barrier (function to protect nerve cells important for brain function from harmful substances), it is unlikely to contribute to the cognitive function improving effect aimed by the present invention. It is possible. On the other hand, since it is pointed out that EPA may be harmful due to excessive intake, it is not necessary to actively mix it with the cognitive function improving agent of the present invention. From such a viewpoint, even if fish oil containing DHA and EPA or several kinds of algae are used as the DHA source, it is preferable to employ one having a low EPA mixing amount. In general functional foods containing DHA, there is a tendency to increase the ratio of EPA by expecting the effect of improving blood flow of EPA and increasing the amount of EPA mixed. The invention has a unique configuration.

[Astaxanthin]
In the present invention, astaxanthin means a product derived from a natural product or obtained by synthesis. Examples of those derived from natural products include crustacean shells such as shrimp, krill and crabs, eggs and organs, various seafood skins and eggs, algae such as the green alga Hematococcus, yeasts such as red yeast Phaffia, Examples thereof include those obtained from seed plants such as marine bacteria, Fukujukusa, and Golden phoenix flowers. Natural extracts and chemically synthesized products are commercially available and are readily available.

  Astaxanthin can be obtained, for example, by culturing red yeast Phaffia, green alga Hematococcus, marine bacteria, etc. in an appropriate medium according to a known method. The green alga Hematococcus is most suitable because it is easy to culture and extract, contains astaxanthin at the highest concentration, and has high productivity.

  Astaxanthin is used as an extract of astaxanthin obtained by the above method and a powder or aqueous solution containing them, or dried products such as red yeast Phaffia, green alga hematococcus, marine bacteria, and crushed products thereof. it can.

“Weight ratio of DHA: Astaxanthin”
The cognitive function improving agent of the present invention contains docosahexaenoic acid (DHA) and astaxanthin. This cognitive function improving agent preferably contains docosahexaenoic acid (DHA) and astaxanthin in a weight ratio (DHA: astaxanthin) of 1000: 1 to 1: 1, and is 1000: 1 to 50: 1. Is more preferable. If it is less than this weight ratio, the effect of astaxanthin on DHA cannot be expected, and even if it contains a large amount of astaxanthin exceeding a certain weight ratio, the effect on the effect as the cognitive function improving agent of the present invention is small. Conceivable. As for the weight ratio, it is more preferable that there is more astaxanthin ratio than 800: 1. Moreover, it is more preferable that there is more DHA ratio than 100: 1, and it is more preferable that there are more DHA ratios than 200: 1. By mixing at such a weight ratio, an effective function can be obtained even with a low DHA content. For mixing DHA and astaxanthin, powders, liquids, oils, and the like containing the respective components may be mixed according to the amount of the components.

  The cognitive function improving agent of the present invention further includes at least one component selected from the group consisting of arachidonic acid, lycopene, phosphatidylcholine, phosphatidylserine, ginkgo biloba extract, niacin, pantetonic acid, and various vitamins. It is preferable to contain. These components are components that are expected to affect the nervous system, and can exhibit a synergistic effect with the combination of DHA and astaxanthin of the present invention to enhance the cognitive function improving effect. Alternatively, even if the DHA amount is set to a low value, it is easy to obtain the effect stably.

"Formulation"
The cognitive function improving agent of the present invention can be ingested in the form of a solid preparation such as a capsule such as a soft capsule, a hard capsule or a tablet. In addition to this, it may be prepared and taken in a dosage form of a preparation suitable for oral administration such as jelly or liquid. This preparation is preferably in the form of a solid preparation that is easy to control the daily intake amount orally administered and that can be easily encapsulated to suppress odor and taste such as DHA.

  An example of a preferable formulation form of the cognitive function improving agent of the present invention is a content containing DHA and astaxanthin in the above-described weight ratio, and has a coating mainly composed of gelatin or the like containing this content liquid, and has a short diameter. Soft capsules such as a substantially spherical shape having a length of 6 to 9 mm and a major axis of 12 to 18 mm, a tapered ellipse shape (a shape generally referred to as a drop shape), and a rugby ball shape are exemplified. As the contents of the soft capsule, fish oil or several types of algae-derived fats and oils can be used as DHA, and astaxanthin and other components mixed as appropriate can be suspended and dissolved in the fats and oils. Moreover, you may add a fragrance | flavor, a sweetener, a pigment | dye, an excipient | filler, a stabilizer, a flavoring agent, a diluent, surfactant, an emulsifier, a solubilizer, an absorption promoter, a bulking agent, a preservative, etc.

  Furthermore, the coating film used for soft encapsulation is a capsule film. By sealing the content liquid with this coating film, the content liquid can be maintained in its ability to prevent oxidation and the flavor of the content liquid. It can be formed as a capsule that is difficult to perceive. The coating is mainly gelatin and is preferably a soft capsule. In some cases, glycerin, sorbitol, a pigment, or the like is appropriately added to the coating, and a solvent such as water is added to facilitate the molding process. In this molding, a capsule is generally prepared by simultaneously filling a film with a coating such as gelatin and molding the film and forming the film. Examples of this forming method include a rotary method and a double nozzle method. In particular, soft capsules are easier to swallow than powders and granules because of their high specific gravity.

  The cognitive function improving agent is preferably used so that it is orally administered as a capsule tablet containing 30 mg to 200 mg of DHA per tablet. For details, as will be described later, the cognitive function improving agent of the present invention is characterized in that 200 mg or more and 600 mg of docosahexaenoic acid per day is used orally. At this time, in order to control the intake by this amount, when the intake is controlled using tablets containing 30 mg or more and 200 mg or less of DHA per tablet, the number of oral administrations per day, that is, the amount to be consumed is ingested. It is easy to manage as an amount that can reduce the swallowing burden on the person and prevent excessive intake.

“Intake (oral dose)”
The cognitive function improving agent of the present invention is used so that 200 mg or more and 600 mg of DHA per day is orally administered to humans.
Such an oral dose is a dose that has not been considered effective in the conventional way of thinking. On the other hand, since it is an amount that is sufficiently smaller than the intake (for example, 2 g / day as the FDA recommended value) that has been suggested to have a negative effect on health, it can be managed as an intake that is excellent in terms of safety. it can.
As described above, the amount to be administered orally is determined by determining the dose of DHA per day (intake amount) for humans. Since the cognitive function-improving agent of the present invention is continuously consumed in daily life, it is generally used as the total intake amount of adults rather than managing it strictly as the intake amount per body weight.

  This amount is the amount taken as the cognitive function improving agent of the present invention. In other words, in the present invention, DHA and astaxanthin are mixed and orally administered as appropriately formulated into a solid, jelly, or liquid form, and the oral dose is controlled by the effect of this mixing It is. Therefore, this intake is an amount taken from what was prepared as a mixture, for example, a setting as an amount taken separately from a normal meal as a capsule etc. Since DHA that is simply ingested from a diet is not mixed with astaxanthin, it is considered that there is little influence on the present invention by ingesting them separately.

"Cognitive function improving agent"
The cognitive function improving agent of the present invention improves human cognitive function. The cognitive function indicates that the current situation is grasped and the best action is selected based on past experience and knowledge. Its function is thought to be mainly played by the limbic cortex of the cerebrum. DHA is thought to affect neurotransmitters, activities of membrane-bound enzymes and ion channels, gene expression, synapses, etc. in the brain, but the mechanism of action is not yet clear There are many different findings depending on the specific intake. The cognitive function improving agent of the present invention is considered to be effective in a wide generation of teenagers to 80s, especially from students who need cognitive function to elderly people who are declining in cognitive function. This cognitive function can be evaluated by, for example, a Stroop test or a revermede action memory test (a test method adopted for memory failure diagnosis). As expressions almost synonymous with this cognitive function improvement, expressions such as judgment power improvement, judgment ability improvement, judgment ability improvement, learning ability improvement, and the like may be performed.

[Test method]
Study design: A randomized, double-blind, placebo-controlled, parallel group comparison study was conducted.
Ingestion start number of subjects: Ingestion was started with 66 people. (Breakdown: test drug intake group: 33, placebo intake group: 33)
Assignment method: The person in charge of assignment created an assignment table using random numbers and assigned assignment numbers to the test agents. The assignment table was sealed by the person in charge of assignment and kept sealed until the assignment table was opened. After fixing the analysis subject and data, the person in charge of assignment opened the assignment table and disclosed the information. In the event that serious adverse events occur and urgent matters are required, the allocation table will be opened as necessary and only the minimum necessary information will be disclosed, but serious adverse events will occur. Since it did not, the allocation table was not opened halfway.

[Subject selection criteria and exclusion criteria]
Those who met the following selection criteria and did not violate the exclusion criteria were selected as subjects.
・ Criteria for selection (1) Men and women aged between 50 and 70 at the time of obtaining consent (2) Those who have a relatively low intake of n-3 fatty acids derived from normal diets (3) Blood in the pre-examination In addition to this selection criterion, those who have a high constitutional fat level are preferentially selected. In addition to this selection criterion, an exclusion criterion is provided for those who have a constitution or symptom that greatly affects the test. In addition, as described in the selection criterion (2), this group of subjects is a person who consumes a relatively small amount of n-3 total fatty acids, and the amount of intake is the 50th percentile of the national DHA intake. It is considered to be less than 0.260 g / day, and is assumed to be about 0.05 to 0.15 g / day.

[Subject restrictions (prohibited items)]
The investigator and the person in charge of the study instructed the subject to live a normal life before participating in the study and observe certain restrictions during the study period.

[Test agent]
Type and name of test agent (1) Test agent: Agent containing DHA / EPA and astaxanthin and other components (2) Placebo agent: Agent not containing DHA / EPA and containing astaxanthin and other components

  The shape, raw materials, participating components, etc. of the soft capsule (A) used as the test agent and the soft capsule (B) used as the placebo are shown below.

[Molding of soft capsule (A)]
A soft capsule (A) was prepared as a cognitive function improving agent of the present invention. The average concentration of each component per tablet (average weight 470 mg) of this soft capsule (A) is as follows.
DHA: 136mg
EPA: 14.8mg
Astaxanthin: 0.33mg
Vitamin E: 3mg
Lycopene: 0.02mg
Phosphatidylcholine: 0.40mg
Phosphatidylserine: 0.15mg
In addition, in manufacturing a soft capsule, the above-mentioned component is the content liquid of the soft capsule (A) obtained by mixing the above-described components such as astaxanthin (derived from Haematococcus alga pigment) into purified fish oil (derived from tuna and bonito). It is a component contained inside. Moreover, in this soft capsule (A), DHA: astaxanthin was set to 408: 1. However, it is considered that the effect of the combination of DHA and astaxanthin tends to be understood logarithmically due to the effect in the human body. Even if the weight ratio when combining these is changed as a tablet, it is thought that effectiveness is easily maintained.

  Further, the content liquid of this soft capsule (A) was mixed in a ratio of 68 parts by weight of gelatin and 24 parts by weight of glycerin, 7 parts by weight of beeswax, and 1 part by weight of colorant as a solid content. By using a coating sheet molded using a coating solution prepared by diluting with water as described above, a soft capsule having the shape shown in FIGS. 1 and 2 was formed by inclusion by a known rotary method. The thickness of the coating sheet at the time of molding was about 800 μm, and was dried at room temperature after molding to remove excess water. This soft capsule (A) was molded with a major axis b12.4 mm and a minor axis a8.2 mm.

[Molding of soft capsule (B)]
Molding was performed according to the preparation of the soft capsule (A) except that olive oil was used instead of the DHA and EPA source fish oil used in the content liquid of the soft capsule (A) described above.

[Intake of test agent, intake method and intake period]
The test agent was ingested 4 grains a day with a sufficient amount of water or lukewarm water. Even if they forgot to take it, they were instructed not to take too much the next day. The intake period was 12 weeks.

[Rationale for setting the intake of components involved]
The amount of DHA 544 mg / day and EPA 59.2 mg / day in the test agent (soft capsule (A)) 4 capsules per day (reference amount) was set as the intake amount of the test agent.

[Examination schedule]
The person in charge of the study facility commissioned by the study investigator will give an explanation of consent, and the lifestyle candidate questionnaire, interview (physical condition confirmation) and physical measurement (height) will be given as prior 1 to 222 subject candidates who have obtained consent. , Body weight), physical examination, clinical examination, and dietary survey. For 96 subjects who were judged to be qualified in advance 1, a cognitive impairment test using the interview (physical condition confirmation) and the revised Hasegawa simplified intelligence evaluation scale was conducted as advance 2. Conducted a medical examination (physical condition check) and a brain functional test as a pre-intake brain function test for 88 subjects judged to be eligible in advance 2 and a medical examination (physical condition check) as a blood sampling test within 7 days after the brain function test And physical measurements, physical examinations, clinical tests, blood flow measurements, and dietary surveys. From the results of the pre-ingestion test, 66 subjects were selected who met the purpose of the test. The study facility distributed test agents assigned to the subjects and diaries during the intake period. During the intake period, the intake of the test agent and the diary were recorded every day.

  In the 4th week (29th day) and 8th week (57th day) of intake of the test agent, medical examination (physical condition confirmation), physical measurement, physical examination, and clinical examination were carried out as various tests. In the 12th week (85th day), a medical examination (physical condition check) and a brain functional test are performed as a brain function test, and a medical examination (physical condition check), physical measurement, and physical examination are performed as a blood sampling test within 7 days after the brain function test is performed. Clinical tests, blood flow measurements, and dietary surveys were conducted.

[Inspection date]
Tests were performed on a total of 8 visits. The prescribed inspection dates were as follows.
(1) Pre-intake test Blood sampling was performed within 7 days after the brain function test.
(2) Examination of 4th and 8th week of ingestion The prescribed date of inspection for 4th week of ingestion is the 29th day of ingestion, and the prescribed date of 8th ingestion is on the 57th day of ingestion. If the examination was not possible, the examination date was changed within the range of 7 days before and after the prescribed examination date at the discretion of the investigator.
(3) Ingestion Week 12 Examination The ingestion week 12 examination was performed between the ingestion days 85-92. The blood sampling test was performed within 7 days after the brain function test. A blood sampling test was performed at the latest 92 days after the start of ingestion.
(4) Re-examination and follow-up survey Re-examination and follow-up inspection were conducted as necessary.

[Adverse event]
・ Definition of adverse events Any undesired medical event newly or worsened in the subject after taking the test agent (abnormal changes such as subjective symptoms, objective findings, physical examination, clinical examination, etc. that occurred after taking the test agent) ) Was an adverse event and was not related to the causal relationship with the test agent.
・ Judgment of adverse events Adverse events as subjective symptoms and objective findings were determined by the investigator. For abnormal fluctuations (adverse events) in individual test values, the abnormal fluctuation criteria set by the Japanese Society of Chemotherapy (Chemotherapy. 58. 484-493 (2010) ), The investigator determined the adverse event.
・ Procedure for reporting serious adverse events If the observed adverse event is serious, the study investigator should immediately report the occurrence to the head of the study medical institution, Although it was stipulated that a report on serious adverse events was prepared and reported to the head of the studying medical institution and the sponsor, the study sponsor was contacted, but no serious adverse events occurred.

[Brain function test procedure]
(1) Word memory test This test evaluates the capacity of immediate memory and short-time memory, as well as its retention and reproduction ability. After 10 words were heard one by one, they were repeated, and as soon as the 10th word was repeated, these words were recalled (immediate recall). For words that could not be recalled by themselves, the number of words that could be recalled by giving hints of initial sounds was taken as the number of CUE recalls. Number of CUE replays that were instructed to be remembered during self-recalling and CUE-recognized words even during the execution of the interference task (word recall test) and that were replayed with hints of spontaneous replay and initial sound after 5 minutes Was recorded. Different words were memorized in each of the pre-intake brain function test and the 12th week brain function test.
The larger the number of spontaneous recalls and spontaneous regenerations (the higher the numerical value), the higher the memory.

(2) Word recall test This test evaluates memory recall ability, that is, fluency. Fluency is the ability to process and output a large amount of information (mainly linguistic information) appropriately and quickly, and is mainly attributed to the action of the frontal lobe. Teach me to say as many fruit names as you know, and write down what you could enumerate in 60 seconds after the first one was named (Recall Test (1)). The number was recorded excluding duplicates. The same applies to “words beginning with“ a ”” (recall test (2)) and “animal name” (recall test (3)). The same test was conducted at each time of pre-ingestion brain function test and week 12 brain function test. The higher the number of recalls (the higher the number), the higher the recall ability.

(3) Stroop test This test evaluates the ability to identify and process two different pieces of information (language information and color vision information) that simultaneously enter the brain, and can evaluate higher-order brain functions. This test is composed of sub-tests of four steps (steps 1, 2, 3, and 4), and the more complicated information processing ability is required as the steps progress, the degree of difficulty increases step by step.

(4) Japanese version of Livermead Behavior Memory Test Japanese version of Livermead Behavior Memory Test (Reiko Watamori, Hiromi Hara, Takashi Miyamori, et al .: Japanese version of Livermead Behavior Memory Test, Chiba Test Center, 2002. (Wilson BA. , Cockburn JM., Baddeley AD .: The Rivermead Behavioral Memory Test (RBMT). Suffolk; Thames Valley Test Company: 1985.)). There are nine sub-items: first and last name, belongings, promises, pictures, stories, facial photos, directions, requirements, orientation and date, scoring records the raw points for each sub-examination, and standard profile points (0 The screening point (1 point or 0 point) was calculated in terms of ~ 2).
In either case, the higher the value, the higher the memory function.

[Evaluation of effectiveness]
[Effectiveness analysis person]
Among subjects who completed all of the predetermined test schedule and test content, those who excluded subjects who met certain criteria were included in the analysis of effectiveness.

[Effectiveness evaluation items]
(1) Main evaluation items: brain function (word memory test, word recall test, Troop test)
(2) Secondary endpoint: brain function (Japanese Livermead behavioral memory test (screening points, standard profile points))
(3) Evaluation method For each item, the amount of change at each time point before and after ingestion was evaluated by comparison between groups. For each item of brain function, the amount of change before ingestion at the 12th week of ingestion was compared using a two-sample t-test for the placebo agent ingestion group and the test agent ingestion group. In each group, the amount of change from before ingestion at each time point after ingestion was evaluated using a one-sample t-test.

[Numeric display and significance level]
Numerical values are shown as mean ± standard deviation, and the significance level of the test was 5% on both sides.

[result]
[Subject classification and breakdown]
Based on the results of the pre-inspection and pre-intake inspections, 66 subjects (33 in the P group and 33 in the A group) that meet the selection criteria and do not meet the exclusion criteria were selected as eligible subjects. Height, weight, and pulse rate were all within the standard range or normal range.
As a result of excluding 11 persons (P group 6 persons, A group 5 persons) that the investigator judged to have an influence on the analysis of effectiveness before opening the allocation table, the analysis of effectiveness was 54 persons (26 persons in P group) , Group A 28 people). The safety evaluation was based on 66 subjects (33 in the P group and 33 in the A group) who took the test agent at least once during the test period for adverse events. The subjects were 65 persons who completed the above (32 persons in the P group, 33 persons in the A group). The intake rate of the test agent was 86.8 to 101.1%. A summary of the background factors of this test subject is shown in a table in FIG.

[Subject background factors]
The comparison between the two groups was performed by χ ² test for gender and 2-sample t test for other items. The average age of group P was 59.1 years (15 men, 11 women), and the average age of group A was 57.4 years (13 men, 15 women). In addition, none of the physical measurement values, physical examination values, n-3 fatty acid intake, and blood triglyceride values showed any imbalance among the groups.

[Evaluation of effectiveness]
[Main endpoint: brain function (word memory test)]
The number of immediate spontaneous recall, the number of immediate spontaneous recall + CUE recall, the number of spontaneous regenerations after 5 minutes, the number of spontaneous regenerations after 5 minutes + the number of CUE regenerations, and the transition of measured values and the amount of change from before intake are shown in FIG. .
(1) Immediate number of spontaneous recalls Before ingestion, it was 5.2 ± 1.6 in the A group and 4.8 ± 1.3 in the P group, and there was no significant difference between the groups. The amount of change before and after ingestion was 0.3 ± 2.0 in the A group and 0.7 ± 1.6 in the P group, and there was no significant difference between the groups. Moreover, the significant increase of the score was recognized by the P group compared with before intake (p <0.05).
(2) Number of immediate spontaneous recalls + number of CUE recalls Before ingestion, there were 2.8 ± 1.5 in the A group and 3.2 ± 1.6 in the P group, and there was no significant difference between the groups. The amount of change before and after ingestion was −0.1 ± 2.1 in the A group and 0.7 ± 1.7 in the P group, and there was no significant difference between the groups. Moreover, the significant fall of the score was recognized by the P group compared with before ingestion (p <0.05).
(3) Number of spontaneous regeneration after 5 minutes Before ingestion, it was 4.9 ± 2.3 in the A group and 4.2 ± 1.8 in the P group, and there was no significant difference between the groups. The amount of change before and after ingestion was −0.2 ± 2.6 in the A group and −0.2 ± 2.3 in the P group, and there was no significant difference between the groups. There was no significant change in the comparison before and after intake.
(4) Number of spontaneous regeneration after 5 minutes + number of CUE regeneration Before ingestion, it was 2.5 ± 1.6 in the A group and 2.5 ± 1.1 in the P group, and there was no significant difference between the groups It was. The amount of change before and after ingestion was −0.5 ± 2.1 in the A group and −0.3 ± 1.7 in the P group, and there was no significant difference between the groups. There was no significant change in the comparison before and after intake.

[Main endpoint: brain function (word recall test)]
For the recall test (1), the recall test (2), and the recall test (3), the transition of measured values and the amount of change from before ingestion are shown in FIG.
(1) Recall test (1)
Before ingestion, there were 13.3 ± 3.0 in the A group and 12.5 ± 2.6 in the P group, and there was no significant difference between the groups. The amount of change before and after ingestion was −0.4 ± 2.5 in the A group and 0.2 ± 2.9 in the P group, and there was no significant difference between the groups. There was no significant change in the comparison before and after intake.
(2) Recall test (2)
Before ingestion, there were 12.7 ± 4.1 in group A and 11.8 ± 4.7 in group P, and there was no significant difference between groups. The amount of change before and after ingestion was 1.4 ± 2.3 in the A group and 2.0 ± 3.4 in the P group, and there was no significant difference between the groups. In addition, a significant increase in scores was observed in both groups compared to before intake (p <0.01).
(3) Recall test (3)
Before ingestion, there were 17.7 ± 4.7 in the A group and 18.0 ± 4.8 in the P group, and there was no significant difference between the groups. The amount of change before and after the intake was 0.9 ± 3.4 in the A group and 1.2 ± 2.9 in the P group, and there was no significant difference between the groups. There was no significant change in the comparison before and after intake.

[Main endpoint: brain function (Stroop test)]
FIG. 6 shows the amount of change for the number of seconds in step 1, the number of seconds in step 2, the number of errors and seconds in step 3, and the number of errors and seconds in step 4. The number of errors in Steps 1 and 2 was omitted because the amount of change was 0 in both groups.

(1) Step 1
The number of errors was 0.0 ± 0.0 in the A group and 0.0 ± 0.2 in the P group before ingestion, and there was no significant difference between the groups. There was no change before and after intake in both groups.
The number of seconds was 47.0 ± 10.7 seconds in the A group and 48.9 ± 14.6 seconds in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was −4.5 ± 7.2 seconds for the A group and −6.4 ± 12.1 seconds for the P group, and there was no significant difference between the groups. In addition, a significant decrease in the number of seconds was observed in both groups compared to before intake (Group A: p <0.01, Group P: p <0.05).
(2) Step 2
The number of errors was 0.0 ± 0.2 in the A group and 0.0 ± 0.0 in the P group before ingestion, and there was no significant difference between the groups. There was no change before and after intake in both groups.
The number of seconds was 45.3 ± 10.1 seconds in the A group and 46.1 ± 9.2 seconds in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was −3.6 ± 6.0 seconds for the A group and −4.7 ± 7.7 seconds for the P group, and there was no significant difference between the groups. In addition, a significant decrease in the number of seconds was observed in both groups compared to before intake (p <0.01).
(3) Step 3
The number of errors was 0.1 ± 0.4 in the A group and 0.0 ± 0.0 in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was −0.1 ± 0.4 in the A group and 0.2 ± 0.5 in the P group, and the amount of decrease in the A group was significantly larger than that in the P group. (P <0.05). In addition, a significant increase in the number of errors was observed in the P group compared to before intake (p <0.05).
The number of seconds was 49.9 ± 17.9 seconds in the A group and 45.6 ± 7.8 seconds in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was −5.2 ± 11.0 seconds for the A group and −1.0 ± 6.8 seconds for the P group, and there was no significant difference between the groups. In addition, a significant decrease in the number of seconds was observed in group A compared to before ingestion (p <0.05).
(4) Step 4
The number of errors was 0.2 ± 0.5 in the A group and 0.1 ± 0.3 in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was 0.2 ± 1.4 in the A group and 0.0 ± 0.4 in the P group, and there was no significant difference between the groups. There was no significant change in the comparison before and after intake.
The number of seconds was 57.6 ± 11.0 seconds in the A group and 59.0 ± 6.6 seconds in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was −2.6 ± 10.2 seconds for the A group and −2.7 ± 9.4 seconds for the P group, and there was no significant difference between the groups. There was no significant change in the comparison before and after intake.

[Secondary evaluation item: Brain function (Japanese version of Livermead Action Memory Test)]
Regarding standard profile points and screening points, the transition of measured values and the amount of change from before intake are shown in FIG.
(1) Standard profile point Before ingestion, the A group was 20.6 ± 2.2 points, the P group was 19.1 ± 2.9 points, and the A group was significantly higher than the P group. The amount of change before and after ingestion was 0.6 ± 2.7 points in the A group and 1.9 ± 2.9 points in the P group, and there was no significant difference between the groups. Moreover, the significant increase of the score was recognized by the P group compared with before intake (p <0.01).
(2) Screening points Before ingestion, the A group was 9.7 ± 1.3 points, the P group was 8.7 ± 1.9 points, and the A group was significantly higher than the P group. The amount of change before and after ingestion was 0.3 ± 1.8 points in Group A and 1.0 ± 2.1 points in Group P, and there was no significant difference between groups. Moreover, the significant increase of the score was recognized by the P group compared with before intake (p <0.05).

[Layered analysis]
For the Japanese version of the Livermead Behavior Memory Test, the standard profile score and screening score each have a cut-off score according to their age, and before and after each score is a criterion for determining whether or not memory impairment is suspected. Since cut-off scores differ between 40-59 years old and over 60 years old, the Japanese version of the Livermead Behavior Memory Test was evaluated for the 50-year-old subject group and 60-year-old subject group.
The subject background factors are shown in FIG. Since there was an imbalance between groups for some subjects for each subject group, details will be described below.
In subjects in their 50s, the P group was significantly higher than the A group in terms of systolic blood pressure. Although there was no significant difference between groups, the intake of n-3 fatty acids was 2.01 ± 0.52 g / day for group P and 1.79 ± 0.37 g / day for group A. Is high (p = 0.192), blood triglyceride level is 127.0 ± 42.4 mg / dL in group P, 152.9 ± 67.0 mg / dL in group A, and high in group A (P = 0.245). In the 60-year-old subject group, the A group was significantly higher than the P group in terms of systolic blood pressure and pulse rate. Moreover, about the n-3 type | system | group fatty acid intake, the P group was 1.73 +/- 0.43g / day, the A group was 2.21 +/- 0.53g / day, and the A group was a significantly high value. Regarding blood triglyceride levels, there was no significant difference between groups, but P group was 142.5 ± 51.5 mg / dL, A group was 118.2 ± 29.0 mg / dL, and P group was The price was high (p = 0.193).

FIG. 9 shows the transition of the measured value of the Japanese version of the Livermead Behavior Memory Test and the amount of change from before ingestion.
(1) Standard profile points For subjects in their 50s, before ingestion, it was 21.1 ± 2.1 points in Group A and 19.0 ± 3.3 points in Group P, and Group A compared to Group P Was significantly higher. The amount of change before and after ingestion was 0.2 ± 2.5 points in the A group and 2.4 ± 2.9 points in the P group, and the P group was significantly larger than the A group (p <0.05). ). Moreover, the significant increase of the score was recognized by the P group compared with before intake (p <0.05).
Regarding the 60-year-old subject group, before ingestion, it was 19.9 ± 2.3 points in the A group and 19.1 ± 2.7 points in the P group, and there was no significant difference between the groups. The amount of change before and after ingestion was 1.5 ± 2.8 points for Group A and 1.4 ± 3.0 points for Group P, and there was no significant difference between the groups. There was no significant change in comparison before and after intake.
(2) Screening points For subjects in their 50s, 9.9 ± 1.2 points in the A group and 8.6 ± 2.0 points in the P group before ingestion. Significantly higher. The amount of change before and after ingestion was −0.1 ± 1.8 points in the A group and 1.5 ± 2.0 points in the P group, and the P group was significantly larger than the A group (p <0. 05). Moreover, the significant increase of the score was recognized by the P group compared with before intake (p <0.05).
For the 60-year-old subjects, 9.2 ± 1.4 points in the A group and 8.7 ± 1.9 points in the P group before ingestion, and there was no significant difference between the groups. The amount of change before and after ingestion was 1.1 ± 1.5 points for Group A and 0.6 ± 2.1 points for Group P, and there was no significant difference between the groups. Moreover, the significant increase of the score was recognized by the A group compared with before intake (p <0.05).

[Evaluation of safety]
[Adverse event]
It was concluded that there were no adverse events (secondary effects) attributable to the test agent.
[Measured value / inspection value fluctuation]
Regarding the changes in hematological tests, blood biochemical tests, and urinalysis, the association with the test agent was determined to be “none”.

[Discussion]
In this study, for men and women between the ages of 50 and 70 years of age, an agent containing DHA and EPA (DHA / EPA-containing agent) was ingested continuously for 12 weeks, and the effect on brain function was randomized, double-blind Examination was performed by comparison between the placebo-controlled parallel groups.

  As a result, the “Step 3 error” in the Stroop test was significantly increased in the placebo-administered group, but slightly decreased in the test-agent-ingested group, and a significant difference between the groups was observed in the amount of change. The Stroop test is a test for evaluating the ability to identify and process two different information (language information and color vision information) that enter the brain simultaneously, and is used as an evaluation test for prefrontal function (cognitive function). Since this test is a test for evaluating brain function for a period of 3 months and a relatively short period, the difference may become more noticeable when taken for a longer period.

  As a brain function test, a word memory test and a recall test were also performed, but no significant differences between groups were observed. The memory function is said to be composed of three stages: “memorization: memorizing information”, “retention: retaining memorized information”, and “reproduction: reproducing information” (Yoko Imamura) , Clinical Higher Level Brain Function Evaluation Manual 2000, Second Revised Edition, Shinsei Medical Publishers, 2001). In the test of memory ability evaluation, improvement by the practice effect was seen, but in this test, the number of immediate spontaneous recalls increased in both groups, and the average score in both groups was 5.5 in both groups. Other than the number of immediate spontaneous recalls, the scores of other words in the memory test and the word recall test are the same in the 12th week. It was speculated that there was no difference.

  The Japanese version of the Livermead Behavioral Memory Test is used as an evaluation test for general memory and cognitive impairment, and is characterized by measuring memory under conditions similar to daily life. Four types of parallel tests with the same level of difficulty have been prepared, and the contents of the tests can be used to continuously evaluate memory impairment by eliminating practice effects. The screening score increased significantly, and the practice effect could not be completely eliminated. The scores at the 12th week were almost the same in both groups, and it was considered that the increase in scores reached the upper limit, similar to the word memory test and the recall test.

  The Japanese version of the Livermead Behavior Memory Test shows a cut-off score that is a guideline for determining the possibility of memory impairment at 40 to 59 years old or younger and 60 years old or older. Since the effects may change depending on the age, we conducted a stratified analysis in the 50s and 60s and evaluated the Japanese version of the Livermead Behavior Memory Test.

  As a result, in the 60-year-old subject group, the screening score increased significantly in the DHA / EPA-containing agent intake group. Regarding the measured values at 12 weeks, the DHA / EPA containing group increased in both standard profile points and screening points to the same level as those in the 50s, and exceeded the practice effect compared to the placebo group. Showed an increase. In relation to the increase in scores at older ages, regarding the absorbability of DHA / EPA-containing agents, the higher the age, the higher the DHA concentration in triglycerides and the higher the DHA concentration in phospholipids. It is shown that an increasing trend was observed.

  The DHA / EPA-containing agent used in this test contains astaxanthin, which is one of carotenoids having strong antioxidant power. For this reason, it is considered that the test agent has a higher protective effect of DHA due to its antioxidant action than ingesting DHA alone. Since the antioxidant activity in the body decreases with aging, it has been considered that the antioxidant activity of astaxanthin appears more prominently in older test subjects.

  In this study, scores were improved in the 60-year-old subjects, although the intake was about ½ compared to the amount of DHA / EPA taken in the previously reported brain function test. The reason for this is that the absorbability of DHA is increased by the inclusion of astaxanthin. In the 50-year-old subject group, a significant difference between groups was observed for both standard profile points and screening points. About this, the standard profile point before ingestion of the placebo drug intake group, and the screening point are significantly lower than the test drug intake group, and both groups have reached the upper limit of the increase in score, apparently, This was considered to have improved in the placebo group.

  Regarding safety, an adverse event was observed in 1 person in the P group, and peri-ocular mild redness (no fatigue) was determined by the investigator to be “possibly none” with the test agent. The incidence of adverse events was 18.2% in group P and 42.4% in group A, which was higher in group A. However, it was not a particularly high rate compared to other studies, but individual adverse events. In all of these studies, it was judged that there was no relationship with the test agent, and it was considered that there was no problem in safety. In the blood test of the placebo group, the platelet count, Na, K, systolic blood pressure, diastolic blood pressure were significantly increased, and ALT (GPT), γ-GT (γ-GTP), Na, and CL were significant. A decrease was seen. In the DHA / EPA containing group, albumin, creatinine, total cholesterol, AST (GOT), ALT (GPT), Na, K, body weight, BMI, systolic blood pressure, diastolic blood pressure significantly increased, CL, blood glucose Although a significant decrease was observed, all were minor fluctuations, and the study investigator determined that there were no clinical problems. From the above, it was concluded that the DHA / EPA-containing agent can be safely ingested under the test conditions.

  If the cognitive function improving agent of the present invention is used, human cognitive function can be improved. Furthermore, it is possible to reduce the risk of excessive intake and continue safely.

1 Contents 2 Peel 10 Soft capsule

Claims (3)

A cognitive function improving agent containing docosahexaenoic acid (DHA) and astaxanthin,
The cognitive function improving agent is used such that 200 mg or more and 600 mg or less of DHA per day is orally administered to a human.   The cognitive function improving agent contains eicosapentaenoic acid (EPA), and the weight ratio (DHA: EPA) of DHA and EPA contained in the cognitive function improving agent is 20: 1 to 5: 1. The cognitive function improving agent according to claim 1, wherein   The cognitive function improving agent according to claim 1 or 2, wherein the cognitive function improving agent is a tablet containing 30 mg to 200 mg of DHA per tablet.