JP6754394B2 - An edible PLs composition and a food composition containing the same to ensure that there is no forgetfulness related to the language and situation of healthy subjects. - Google Patents

Description

The present invention is a safe and stable plasmalogen composition derived from a chicken and for improving cognitive function containing the same, that is, specifically, there is no forgetfulness related to the language and situation of a healthy subject. It is about a food composition for so on. More specifically, the present invention develops a plasmalogen composition having an effect of improving cognitive function (hereinafter, may be referred to as "PLs") and a food for improving cognitive function containing the same. Is intended. The biggest challenge is to demonstrate the effect of improving the cognitive functionality of the plasmalogen-containing food on healthy subjects. In the past, clinical trials have been conducted exclusively to demonstrate the effectiveness of plasmalogen in patients with dementia, and the manifestation of its cognitive function improving effect is still limited, and it is still at the practical level. The reality is that it is far from that (Non-Patent Documents 10 and 11 etc.).

In recent years, plasmalogen has been attracting attention as a therapeutic and / or onset suppression and preventive agent for neurodegenerative diseases and psychiatric diseases (for example, [Patent Document 1]). Plasmalogen is a special phospholipid that is resident in vivo and has a vinyl ether bond to SN-1, and therefore has a specificity that exhibits reducing property. Although this plasmalogen is specific, it accounts for 18% by mass of all phospholipids in the living body, and it is no exaggeration to say that it is one of the versatile phospholipids because of its content.

This plasmalogen is functionally special in that it exerts an important reducing action in the living body, but quantitatively, its existence position is localized to an important membrane in the living body, and various membranes are used. Since it directly protects against oxidative wear and tear, it is not only versatile, but it has become clear that it exhibits multifaceted functions, especially in the brain. Is to be collected. However, as is clear from its molecular structure, plasmalogen is easily oxidized by reducing inside out (radical scavenger property, high radical scavenging sensitivity) and highly hydrolyzable by hydration under acidic conditions (phospholipid). Decomposed into radicals and aliphatic aldehydes). This is considered to be one of the causes hindering the practical organic synthesis of plasmalogen.

Currently, plasmalogens rely exclusively on extraction and purification from living tissues (eg, [Patent Document 2], [Patent Document 3], [Patent Document 4], etc.). In the living body, in view of its importance, it is stabilized by various purposeful mechanisms and is supplemented by the biosynthetic system, that is, the production of endoplasmic reticulum from peroxisomes.

Poultry, especially adult spawning (also referred to as egg collection) chickens, are the mainstream as raw materials for extracting plasmalogen (for example, [Patent Document 1], [Patent Document 2], [Patent Document 3], [Patent Document 4]. ], Etc.), sea squirt-derived offal ([Patent Document 5], etc.) and cultured scallop offal (for example, [Patent Document 6]) have begun to be used. In farmed bivalves, concerns about shellfish poisoning, especially cadmium, that accumulates in the environment (safety concerns) cannot be dispelled.

It is judged that the most rational method for obtaining a safe, inexpensive and stable plasmalogen is through biometabolism. At that time, the selection of the living body is important. Adult laying hens that have already been put to practical use of DHA hen eggs, especially abandoned hens that lay eggs (spawning chickens that are thinned out due to a decrease in spawning efficiency), and more preferably forced hens (regenerate lost feathers by short-term fasting) An example is an egg-laying abandoned chicken by a poultry farming method) that improves egg-laying efficiency. The spawning chicken is literally a poultry whose purpose is to produce chicken eggs, and even the same chicken is a poultry different from a broiler for the purpose of collecting meat. Due to the distribution of eggs, spawning poultry is about 1/10 the size of broilers (10 million to tens of millions) and is bred in the vicinity of the consumption area.

Normally, this spawning poultry farm lays 300 or more eggs per year, but when the age at which spawning efficiency drops is about 700 days, it is thinned out as abandoned chickens, and it is located near the poultry farm. Slaughtered, dismantled and meat-collected chickens are slaughtered and dismantled at a private waste chicken processing center (there are about 40 locations nationwide and the processing capacity is millions to ten millions). The processed products are used for food, but most of them are distributed as low-priced chicken raw materials for processing in the form of minced meat or the like, except for some. Spawning poultry has been forced to sell at a low price due to price competition with bulk imported frozen chicken, but it has been boosted by consumers' safety consciousness in recent years, and it has added value as domestic chicken, and the price is on the rise. However, economically, it cannot compete with the vertically integrated poultry farming of tens of millions of meat-only breeds, and its quality cannot compete with the texture and taste of young chickens (young chickens) that are only 50 days old. The profitability of the spawning and poultry industry continues to be sluggish.

In the above situation, it should be noted that the average annual number of spawning abandoned chickens is 90 million, and the poultry farm delivery price is generally 0 yen, which is cheap. There is. Further, the degree of accumulation of the occurrence is high, and the chickens are carried to the waste chicken processing center as live chickens to obtain a high freshness processed product comparable to that of a broiler.

The present invention is considered to be a globally versatile intractable disease, and Alzheimer's disease (hereinafter referred to as "AD"), which is regarded as an urgent global issue to be held up to the summit ([Non-Patent Document 1]) aiming at its cure. It is intended to throw a stone in the conquest of dementia, including. Recently, most of the chronic neurodegenerative diseases such as AD, Parkinson's disease, amyotrophic lateral sclerosis (hereinafter sometimes referred to as "ALS"), multiple sclerosis, or stroke or head. It has become clear that acute brain damage such as trauma is accompanied by chronic inflammation of the central nerves (brain, spinal cord, etc.).

It is also possible that these diseases are caused and progressed by central inflammation ([Non-Patent Document 2]). For example, it has been reported that central nervous system inflammation may cause neurodegenerative diseases such as Alzheimer's disease and may promote the disease (for example, [Non-Patent Document 2]). In addition, as a result of analyzing a memory disorder model rat created by intraperitoneal administration of LPS (lipopolysaccharide), which is a substance that causes inflammation, accumulation of Aβ (amyloid β) peptide was observed in the brain, and it was an anti-inflammatory agent. It has been reported that a certain sulindac sulpide improved the symptoms ([Non-Patent Document 3]). Furthermore, it has become clear that central nervous system inflammation is frequently observed in mental illnesses such as depression and autism, developmental disorders, and even in the normal aging process.

Based on the above, infection by preventing or treating central nervous system inflammation and related neuronal cell death ([Non-Patent Document 4]), neurodegeneration inhibition ([Patent Document 2]) and Aβ intracerebral accumulation. Prevents nerve cells from being damaged by sexual inflammation and causing neuropathy, or treats neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, or mental illness and development such as schizophrenia, depression, and autism. It is expected to be able to treat disorders ([Non-Patent Document 3]). Under such circumstances, a method capable of effectively treating central nervous system inflammation and the like without side effects is more desired than ever before.

Japanese Patent No. 5847086 Japanese Patent No. 5062873 Japanese Patent No. 5774816 Japanese Patent No. 5430566 JP-A-2007-262024 Japanese Patent No. 5540209 Japanese Patent No. 5360454 Japanese Patent No. 5704493 Japanese Patent No. 5784486

"G8 Dementia Summit" December 11, 2013 London; http://dementiachallenge.dh.gov.uk/2013/12/12g8denetia-summit-agreements/) J. Neuroinflammation, 9: 197, 2012 Ann. N.Y. Acad. Sci., (2012) 1262: 85-92 PLoS ONE.8: e83508.doi: 10.1371 (2013) "Basics and Clinical Practice of Plasmalogen" Sponsored by Plasmalogen Study Group-Effectiveness against Alzheimer's Disease-Symposium Handouts; 2014/02/04 Centennial Hall, Kyushu University School of Medicine Neuroscience Letters, 556 (2013) 104-108) NHK Special Interview Group "Cure Alzheimer's Disease!"-Prescription for the Age of "8 Million People with Dementia"-(2014) p35-51, p76-77, p79-83 (Shufu to Seikatsusha) UMIN R000028474 Evaluation of improvement of brain function by ingestion of plasmalogen 1st International Plasmalogen Symposium; November 7-8, 2016, Kyushu University, Abstracts of Lectures P26 Efficacy and Safety of Plasmalogen Extracted from Scallop in Patients with Moderate to sever Alzuheimaer's Disease: Open Trial 1st International Plasmalogen Symposium; November 7-8, 2016 Kyushu University, Abstracts P24 Efficacy and Safety of Plasmalogen Extracted from Scallop in Patients with Mild Alzuheimaer's Disease and Mild Cognitive Impairment: a Randamized, Double-Blinded, Placebo-Controlled Trial 1st International Plasmalogen Symposium; November 7-8, 2016 Kyushu University, Lecture Abstracts P26 Dement Geriatr Disord Extra 2. 298-303 (2012) Erythrocyte plasmalogen decreased in patients with Alzheimer's disease

The present invention provides a safe and stable plasmalogen composition, and clinical trials of carefully selected healthy subjects in a randomized, double-blind, placebo-controlled manner in which the plasmalogen composition improves the cognitive function. The challenge is to provide safe, stable, and inexpensive foods for improving cognitive function that contribute to the suppression and prevention of neurodegenerative diseases such as dementia and psychiatric disorders.

Surprisingly, the present inventors have found a composition ratio specific to plasmalogen derived from chicken peeled breast, and further researched this as a complex lipid derived from chicken peeled breast. The present invention has been completed by clarifying in clinical studies that the overall cost-effectiveness of human cognitive function can be significantly improved by appropriately blending edible complex lipids). .. The results of the clinical trial are entitled IMPROVEMENT IN COGNITIVE FUNCTION BY SUPPLEMENT CONTAINED PLASMALOGEN FOR HEALTHY JAPANESE-A RAMDOMIZED, DOUBLE-BLI N DED, PLASEBO-CONTROLED STUDY-, and are peer-reviewed "Clinical and New Drugs" Vol. It is published in.

That is, the present invention includes, for example, the subjects described in the following sections.
(1) Peeled breast of spawning abandoned chicken by forced molting (hereinafter referred to as "chicken peeled breast") is treated by an ethanol extraction method to contain at least diacylglycerophospholipid and sphingomyelin. A complex lipid containing 10% by mass or more of ethanolamine-type plasmalogen (described as [PL-PE]) and choline-type plasmalogen (described as [PL-PC]) in terms of dry mass (hereinafter referred to as [PL-PC]). , Called "edible complex lipid")
The edible complex lipid is enzymatically treated with phosphorlipase A1 (PLA1) to decompose diacylglycerophospholipids into free fatty acids and lysophospholipids, and these are removed to remove choline type [PL-PC]> ethanolamine type [PL-PE]. An edible plasmalogen having an ethanolamine-type and choline-type plasmalogen content of 20% by mass to 50% by mass in terms of dry mass, which is the plasmalogen composition derived from the peeled breast of chicken (hereinafter, "PLs"). ”) And prepare
The PLs and the edible complex lipid are appropriately mixed to prepare an edible plasmalogen composition containing both the PLs and the edible complex lipid (hereinafter, referred to as "edible PLs composition").
A stock solution for soft capsules or foodstuffs (hereinafter referred to as "edible PLs composition-δ-tocopherol stock solution") in which the edible PLs composition is uniformly mixed in δ-tocopherol having a mass of 100 to 200 times the mass thereof is used. Formulate,
It was related to the language and situation of a healthy subject containing the edible PLs composition containing the edible PLs composition-δ-tocopherol stock solution and δ-tocopherol containing 100 to 200 mass times the mass thereof as a base. A food composition to help you remember.

Plasmalogen, which is a general-purpose phospholipid resident in the living body, has a history that the living tissue containing the plasmalogen has been conventionally edible. Therefore, the plasmalogen according to the present invention (particularly the plasmalogen extracted from living body, especially chicken tissue) is considered to be extremely safe with almost no concern about side effects. However, once plasmalogen is extracted and separated from living tissue, its stability is lost immediately and the vinyl ether bond is in danger of decomposition. In other words, this means that the [strongly reducing] vinyl ether bond is strongly [easy to oxidize] (= "easy" oxidative decomposition), and some appropriate protective measures must be taken to make effective use of this. It is essential to apply.

In the present invention, as a result of diligent research, a measure for rational breakthrough of the "antinomy" is
1) Purified PLs are appropriately blended with complex lipids derived from chicken breast and having a specific composition ratio.
2) α-tocopherol is used as a solvent for PLs composition,
3) Add δ-tocopherol as an antioxidant,
4) Filled in enteric and high barrier soft capsules,
By doing so, I succeeded in clearing this.

Secure stabilized plasmalogen of the present invention includes the plug Zumaro Gen, neurodegenerative disorders (dementia, Alzheimer's disease, Parkinson's disease, depression, and schizophrenia prodrome) relaxation and for the prevention of It can be used as a supplement ([Non-Patent Document 5]).

Plasmalogen is a component that is abundantly contained in living tissues, and living tissues containing plasmalogen have been conventionally provided for food. Therefore, the food for improving cognitive function of the present invention containing plasmalogen (particularly plasmalogen extracted from living tissue) is safe as a supplement for alleviating and preventing brain dysfunction with almost no side effects. Is considered to be an extremely highly functional food.

A three-step question for selecting subjects regarding cognitive function is shown. The sheet for MMSE is shown. A collection of questions for cognitive function diagnosis is shown. The test result of MMSE is shown. The result of the UK test is shown.

The present invention will be described in more detail below.
The present invention relates to a safe and inexpensive plasmalogen, a study of its cognitive function improving action on humans, and a development of a food for improving cognitive function.

Plasmalogen is a type of ether phospholipid, which is a glycerophospholipid having a long-chain alkenyl group mediated by a vinyl ether bond in SN-1, and is a general-purpose type that accounts for 18% by mass of in vivo phospholipid, but has strong reducing property. It is a general term for special phospholipids that have. This peculiarity imparts remarkable oxidative decomposition and hydrolyzability to plasmalogen, and since it is a typical "antinomy" event in practical use, a rational breakthrough measure is required. The present inventors are safe while maximizing the intrinsic functionality of plasmalogen by rationally avoiding the "antinomy" sexual event, that is, by avoiding the "turning the corner and killing the cow" method. We succeeded in stabilizing the invention and established the present invention.

As raw materials for plasmalogen used in the present invention, chicken meat (particularly breast meat), chicken skin, internal organs (particularly intestine, ovarian fallopian tube gold crown, gizzard), eggs, gala (minced meat preparation by-product), feathers and the like are used. It is preferable to use it. Peeled breast meat, which is generated in a large amount and has a long eating experience, is particularly suitable.

In the present invention, it is particularly preferable to use plasmalogen extracted from chicken tissue (particularly breast meat) as the plasmalogen extracted from living tissue. Among them, chickens, which have been conventionally regarded as edible, are suitable because their safety has been confirmed and stable supply is easy. The method for extracting plasmalogen from living tissue is not particularly limited as long as plasmalogen can be extracted (and purified as necessary), but from the viewpoint of simplicity and cost, extraction and purification are performed as described below. Is preferable. Further, according to the extraction and purification method, diacyl-type glycerophospholipids can be decomposed and removed, which is preferable in that the purity of plasmalogen can be further increased.

Specific examples of the steps of extracting and purifying plasmalogen include the following steps 1) and 2).
1) A step of extracting and fractionating a total lipid fraction (including a low molecular weight water-soluble fraction), a protein fraction, and a neutral lipid fraction from a living tissue into three phases.
Specifically, the following steps 1) to 5) are exemplified.
(1) The process of mincing raw tissue and slowly freezing it.
(2) Frozen minced meat is forcibly thawed, pressed and dehydrated, and then sterilized by high-speed cooking with "overheated water" (Aquagas ^RTM ; [Patent Document 6], [Patent Document 7], [Patent Document 8], [Patent Document 9]). Vacuum high-speed cooling (cooling and dehydration in an oxygen-free atmosphere),
(3) After the above dehydration treatment, a step of adding 3 times (V / W) amount of degassed (deoxidized) ethanol and slowly stirring for 12 hours in a sealed oxygen-free atmosphere to perform extraction.
(4) The process of repeating the above,
(5) A step of coalescing ethanol, distilling off ethanol in an oxygen-free atmosphere, and centrifuging to separate an aqueous layer (water-soluble low molecular weight fraction) to obtain a total lipid fraction.
2) An enzyme is added to the complex lipid fraction to hydrolyze the SN-1 bound fatty acid, the mixed diacylglycerophospholipid is converted into a lyso form, and the plasmalogen is purified by extraction and fractionation with a by-product fatty acid in a hydrophilic solvent. Process.

The extraction is preferably carried out with an organic solvent (for example, one having food suitability such as ethanol, acetone, hexane and the like and at least two or more mixed solvents selected from the group consisting of these) or a hydrous organic solvent. In addition, the chicken tissue to be extracted is preferably treated by the above steps (1) and (2). That is, in order to reduce the amount of ethanol used in the three-phase separation with ethanol in the subsequent step as much as possible, it is required to deoil and dehydrate the living tissue as much as possible at a low temperature and in a short time in an oxygen-free atmosphere.

As for the extraction treatment conditions, in order to minimize the oxidative decomposition and hydrolysis of the contained plasmalogen, it is a short-time stirring treatment at a low temperature in a sealed oxygen-free atmosphere. As a suitable example, ethanol is added to the breast meat of the spawning adult chickens treated in the above-mentioned pretreatments (1) and (2), and the mixture is allowed to stand or slowly agitate at 30 ° C. or higher and 50 ° C. or lower for 180 minutes or longer. The method is exemplified. To 1 kg of the breast meat, add 2 to 4 L of ethanol that has been degassed (deoxygenated) in advance, and leave it to stand under a seal or slowly stir it.

In the hydrous ethanol phase, the ethanol is evaporated and concentrated to separate the aqueous layer, and then hexane that has been degassed in advance is added to extract the phospholipid fraction. The separated aqueous layer and hexane insoluble layer are combined, degassed water is added, and the mixture is allowed to stand at 4 ° C., then centrifuged at a low temperature to remove the insoluble portion, and this is freeze-dried to be low-molecular-weight water-soluble. Obtain a fraction of 12 g. On the other hand, the hexane solution is evaporated to dryness by a conventional method to obtain 5.4 g of a complex lipid fraction.

The phospholipid fraction can be subjected to an enzymatic hydrolysis treatment step to hydrolyze the diacyl-type phospholipid, and plasmalogen can be preferably concentrated. Examples of such a hydrolysis treatment include an enzyme treatment with phospholipase A1 (hereinafter referred to as “PLA1”) ([Patent Document 4]). When treated with PLA1, the mixed diacyl-type glycerophospholipids are decomposed into free fatty acids and lysophospholipids, and these can be extracted and partitioned with acetone and hexane to purify plasmalogen. Removal of free fatty acids and lysophospholipids can be carried out, for example, by partitioning with acetone and hexane.

The origin of PLA1 is not particularly limited as long as the above effects can be obtained, and examples thereof include PLA1 derived from Aspergus oryzae. The PLA1 can be purchased from, for example, Mitsubishi Chemical Foods Co., Ltd. The amount used can be appropriately set according to the amount of complex lipid obtained. Preferably, 0.2 to 200 units / 1 mg of complex lipid can be used, and more preferably, 2 to 200 units / 1 mg of complex lipid can be used. In addition, 1 unit is an amount which changes 1 μmol of a substrate (complex lipid) per minute, and means 1 μmol / min.

The buffer to be used can also be appropriately selected according to PLA1. For example, 0.1 M citric acid-hydrochloric acid buffer (pH 4.5) can be used by dissolving 1 g of the complex lipid in 1 to 30 ml, preferably 5 to 15 ml per 1 g of the complex lipid, and adding a predetermined amount of PLA1. .. As for the reaction conditions, the enzymatic reaction is carried out by stirring in a nitrogen gas atmosphere using a degassed buffer for as short a time as possible, preferably for 1 hour, at a temperature as low as possible, preferably up to 50 ° C.

Avoid enzyme inactivation treatment by heating, cool to room temperature immediately after the reaction is completed, add 2-3 times the amount of hexane that has been degassed in advance, and centrifuge to recover the upper hexane layer. By doing so, the enzyme buffer and the enzyme protein are removed. The hexane solution is partitioned by appropriately adding acetone and water, and further, the solution is partitioned by water or an aqueous solution to remove lysophospholipids and purify the plasmalogen. That is, neutral lipids other than phospholipids are removed with acetone, and plasmalogen and lysophospholipids are separated by water-based solution distribution. The plasmalogen derived from the biological tissue thus obtained can be preferably used as an active ingredient of the supplement for alleviating and preventing the cognitive dysfunction for improving the cognitive function of the present invention.

Complex lipids derived from living tissues and phospholipids of plasmalogens and their composition ratios have unique characteristics depending on the living tissues. The following is an example of the peeled breast of an adult laying hen and the skin, gizzard, intestine, gala and gold crown for comparison.
1. Breast meat
(1) Plasmalogen; [ethanolamine type]: [choline type] = 1: [0.5 to 5]
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [2-10]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [0.5 to 5]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [5 to 50]: 1

2. For comparison 1) Skin (1) Plasmalogen; [Ethanolamine type]: [Choline type] = [1-10]: 1
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [1.5 to 15]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [0.5 to 5]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [1.5 to 10]: 1

2) Gizzard (1) Plasmalogen; [ethanolamine type]: [choline type] = [2 to 15]: 1
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [1-10]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [0.5 to 6]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [1-10]: 1

3) Intestine (1) Plasmalogen; [ethanolamine type]: [choline type] = [2 to 15]: 1
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [2-20]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [1-12]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [1-10]: 1

4) Gala (1) Plasmalogen [ethanolamine type]: [choline type] = [1-10]: 1
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [2-20]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [1-10]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [3 to 25]: 1

5) Gold crown (including ovaries)
(1) Plasmalogen; [ethanolamine type]: [choline type] = 1: [0.0001 to 0.1]
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [1.5 to 15]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [15 to 130]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [20-200]: 1
6) Helmet carcass (peeling)
(1) Plasmalogen; [ethanolamine type]: [choline type] = [0.5-5]: 1
(2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: [2 to 15]
(3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: [0.5 to 5]
(4) [Total glycerophospholipid]: [Total sphingomyelin] = [3 to 20]: 1

The following 1) to 3) are highly specific, and the closest to these are the [helmet carcasses] containing the "vertebra", so to speak, [breast meat] + [bone marrow]. It can be regarded as equivalent to [breast meat], which has a high integrated price-to-performance ratio.
1) Breast meat is [PL-PC]> [PL-PE] and myocardial-like 2) Gold crown is [PL-PE] >>> [PL-PC] and [PL-PC] is almost zero 3) Raw gala Since the incidence is high, the price is less than zero due to industrial waste treatment, the composition is close to [breast meat], and it contains bone marrow (which contains the peripheral tissues of the brain), it has potential added value (brain). Function improvement function) is expected, and it is considered that the integrated price-to-performance ratio is remarkably high.
4) Helmet carcass

The complex lipid and plasmalogen extracted from the living tissue used in the present invention have the contents of ethanolamine plasmalogen and choline plasmalogen as shown in 1) and 2) below in terms of dry mass.
1) The complex lipid is preferably 10% by mass or more.
2) The plasmalogen is preferably 20% by mass to 50% by mass.

The mass ratio and content of ethanolamine plasmalogen and choline plasmalogen can be determined by analyzing a complex lipid or plasmalogen extracted from a living tissue by high performance liquid chromatography (HPLC). Specifically, in HPLC, a chromatogram is obtained by evaporation light scattering detection (ELSD; Evaporating Light Scattering Detector), and the peak area ratios indicating ethanolamine plasmalogen and choline plasmalogen in the chromatogram are obtained. The mass ratio can be calculated. Further, the content can be obtained by calculating what percentage of the peak area of the entire chromatogram the peak area indicating ethanolamine plasmalogen and choline plasmalogen corresponds to.

When the food for improving cognitive function according to the present invention is used as a food additive and / or general food (hereinafter referred to as "various foods"), the various foods may be various plasmalogens themselves. These may be appropriately blended with a base material, a carrier, an additive that is acceptable in terms of food hygiene, and other ingredients / materials that can be used as food. In addition, examples of such various food forms include, but are not limited to, liquid, powder, flake, granule, and paste.

When the food for improving cognitive function according to the present invention is used as a food or drink, the food is plasmalogen, a base material, a carrier, an additive that is acceptable in terms of food hygiene, and other ingredients that can be used as food. The material is appropriately blended (that is, a food composition containing various plasmalogens). For example, processed foods, beverages, health foods (nutritional functional foods, foods for specified health uses, etc.) supplements containing plasmalogens for improving cognitive function or, for example, for preventing and / or alleviating the diseases exemplified above. , Foods for the sick (hospital foods, sick foods, nursing foods, etc.) can be exemplified.

Although not particularly limited, when the various plasmalogens contained in the food are various plasmalogens extracted from the tissues of livestock or poultry (for example, cows, pigs, chickens, etc.), for example, the various plasmalogens are blended. Processed foods such as hamburgers, meat balls, wieners, chicken shavings, chicken skin chips, etc., and health foods (nutritional functional foods, foods for specified health use, etc.) containing processed meat foods, supplements, for the sick It is preferably food or the like. In addition, plasmalogen is made into powder, for example, for beverages (juice, etc.), confectionery (for example, gum, chocolate, candy, biscuits, cookies, okaki, rice cake, pudding, jelly-like confectionery, almond jelly, etc.) It may be contained in various foods and drinks such as tofu), breads, soups (including powdered soups), and processed foods.

When preparing foods and drinks related to the present invention as health foods (foods with nutritional function, foods for specified health use, etc.) and supplements, for example, granules, capsules, tablets (churaburu agents) so as to facilitate continuous ingestion. , Etc.), beverages (drinks, etc.), etc., and among them, capsules, tablets, tablets, jelly, etc. are more preferable from the viewpoint of ease of ingestion, but particularly these. Not limited. The forms of granules, capsules, tablets, jellies and the like can be appropriately prepared according to a conventional method using pharmaceutically and / or food hygiene-acceptable carriers and the like.

The amount of plasmalogen blended in the food or drink according to the present invention is not particularly limited as long as the effect of improving cognitive function can be exhibited, but is preferably 0.00005 to 100% by mass, more preferably 0.0005 to 0.005-. It is 75% by mass, more preferably 0.005 to 50% by mass. The food and drink according to the present invention can be preferably used for improving cognitive function and, for example, for preventing or ameliorating the symptoms of the disease of the above-exemplified sick person.

The hospital food is a meal provided when the patient is admitted to the hospital, the sick person's meal is a meal for the sick, and the care food is a meal for the care recipient. The food and drink according to the present invention is preferably used as a hospital food, a sick person's food, or a long-term care food for a patient who is hospitalized, treated at home, etc. due to the disease of the above-exemplified sick person, or a patient who is receiving nursing care. it can. In addition, it can be taken prophylactically by a person who is highly likely to suffer from the disease of the above-exemplified sick person such as an elderly person.

The target for administering the food for improving cognitive function according to the present invention may be not only humans but also other mammals. Examples of such mammals can be assumed to be raised as livestock or pets, and examples thereof include dogs, cats, cows, horses, pigs, sheep, goats, monkeys, rabbits, mice, rats, and hamsters. ..

The administration time of the food for improving cognitive function according to the present invention is required to be administered, for example, from 5 years before the onset of dementia, more preferably 10 years before, and even more preferably 15 years before the onset of dementia (onset diagnosis). At the present time, it has been clarified that the patient has reached the end stage ([Non-Patent Document 7]). Oral administration is preferable as the preferred administration form in view of the long administration period. The dose of the anti-cognitive function-improving food involved can be appropriately selected according to the age, the degree of symptoms, other conditions, etc. Usually, the amount of plasmalogen in the improved food is preferably 0. It is desirable to use an amount in the range of 001 to 1000 mg, more preferably 0.001 to 100 mg as a guide. It should be administered once a day or in a plurality of times (preferably 2 to 3 times). it can.

The following 1) to 7) are required as requirements for an innovative clinical trial method for predetermined effects (prevention and alleviation of neurodegenerative disorders and psychiatric disorders) of various plasmalogens and their compositions and preparations according to the present invention. It can be illustrated.
1) The subject is characterized by being a healthy person.
According to recent research results in the United States ([Non-Patent Document 7]), the accumulation of Aβ and τ protein in the brain starts at least 10 years, usually 15 to 20 years before the onset diagnosis time. It has been clarified that a kind of "paradigm shift" has been triggered. Early prevention is the decisive factor in suppressing the onset, and this is the reason why it is important to develop the efficacy in healthy to non-diseased patients.
2) It is characterized by using a safe and stable plasmalogen and / or composition and preparation.

Based on the above, the ingredients for the prevention and alleviation of neurodegenerative disorders and psychiatric disorders are required to be ingested for a much longer period of time than before, so that safety, stability and low cost are essential requirements.
3) More preferably, a food for improving cognitive function, which is obtained by adding a safe and stable plasmalogen, is used.
4) The safe and stable plasmalogen is safely extracted from chickens safely bred with a safe feed.
5) It is important to keep the daily intake as low as possible. For example, the range of 0.001 to 1.5 mg is exemplified, and it is characterized by safe and convenient oral administration.

6) Compared to the usual clinical trial period of 1 year or more for the disease subject, it is 2 years or less at the longest, preferably 18 months, more preferably 12 months, preferably 6 months or less and 1 month or more. It is short and can reduce the burden on the subject.
7) Innovative clinical trial method for determining preventive and palliative efficacy of neurodegenerative disease disorder and psychiatric disorder, characterized in that the primary essential test efficacy includes 3 or more items selected from the following groups. Is.

* Uchida / Kleperin test * MMSE
* Resting functional MRI (rs-fMRI) image analysis (Non-Patent Document 6)
* Measurement of PLs content in erythrocytes (Non-Patent Document 12)
* PSOL "Cognitive Function Self-Diagnosis Test" (Non-Patent Document 10)
* RBANS (Non-Patent Document 8)
* Cognitrax (Non-Patent Document 8)
* Wexler memory test (Non-Patent Document 9)

Hereinafter, the present invention will be specifically described based on Preparation Examples and Examples, but the present invention is not limited to the following examples.

Preparation Example 1
[Manufacture of plasmalogen-containing phospholipids extracted from waste chicken skin peeled breast]
1. Preparation of Complex Lipids from Waste Chicken Peeled Breast Fresh peeled breast of spawning waste chicken, which is a chicken tissue, was procured from a waste chicken processing center, and 1 kg of minced meat of about 8 mm was prepared. The minced meat was slowly frozen and stored. Before use, it was forcibly thawed with warm running water and then squeezed to dehydrate and deoil. This was cooked in [overheated water] (aqua gas; RTM) in an “anoxic atmosphere” (oxygen 0.5% by volume or less) for 5 minutes, and then rapidly cooled and dehydrated by a vacuum cooling method. The obtained dehydrated, deoiled, cooked and sterilized breast meat was pulverized at low temperature and then subjected to an ethanol three-phase separation step.

[Three-phase separation step]
The cooked and sterilized crushed breast obtained above was placed in a container and degassed, 800 ml of degassed ethanol was added to this container, and the mixture was sealed and slowly stirred at 35 ° C. for 10 hours. Later, the mixture was allowed to stand under ice-cooling to separate the chicken oil content and the solid-precipitated protein content in the upper layer to obtain a hydrous ethanol fraction (sealed cold storage). 800 ml of degassed ethanol was added to the protein content, and the same extraction operation was repeated. The ethanol phase was separated by centrifugation, ice-cooled under a seal, and concentrated under reduced pressure. The solid content of the water-soluble low molecular weight fraction was filtered off, and then evaporated to dryness under reduced pressure to obtain 7 g of plasmalogen-containing phospholipid (complex lipid).

(1) Fractionation of total lipids The total lipids extracted, distributed and separated by the Folch method (hereinafter, synonymous with “simple lipids + complex lipids”) were 1.8% by mass.
(2) Profile test of phospholipids Measured by the HPLC / ELSD method of the conventional methods [Patent Document 1].
(3) The list notation of the above results in [mg / 100 g breast meat] was as follows.
TL (total lipid): 1,800 , TPL (total phospholipid): 700
PL-PE (ethanolamine type plasmalogen): 61, PE (phosphatidylethanolamine): 44, PL-PC (choline type plasmalogen): 124, PC (phosphatidylcholine): 276, SM (sphingomyelin) ): 32

(4) Composition ratio of peeled breast-specific phospholipids 1) Plasmalogen; [ethanolamine type]: [choline type] = 1: 2
2) Diacylglycerophospholipids; [ethanolamine type]: [choline type] = 1: 6.3
3) [Ether glycerophospholipid]: [Diacyl glycerophospholipid] = 1: 1.7
4) [Total glycerophospholipids]: [Total sphingomyelin] = 16: 1

Preparation Example 2
[Preparation of purified plasmalogen from complex lipids]
1. Preparation of Purified Plasmalogen from Complex Lipid of Waste Chicken Peeled Breast 20 g of the above plasmalogen-containing phospholipid (complex lipid) was added to 400 ml (10 mg / ml 0.1 M citric acid-hydrochloride buffer) of phospholipase A1 (Mitsubishi Chemical Foods) solution. ), And stirred at 50 ° C. for 2 hours under filling with nitrogen gas. Under ice-cooling, double volume of hexane was added, and the mixture was stirred and distributed twice, and the upper layer was recovered and concentrated to dryness. Further, 60 ml of acetone was added to the dry matter, the mixture was stirred and centrifuged, and the operation of collecting the precipitate was repeated twice.

Further, 60 ml of hexane / acetone (7: 3) was added to the precipitate, and the mixture was stirred and centrifuged to recover the liquid layer. After concentrating and drying this liquid layer, 240 ml of hexane / acetone (1: 1) was added and transferred to a separating funnel, 36 ml of water was added, and the mixture was stirred and distributed. The lower layer was discarded, 96 ml of acetone / water (5: 3) was added to the upper layer, and the mixture was stirred and distributed. The upper layer was recovered and quickly dried under reduced pressure to obtain purified plasmalogen derived from chicken skin peeled breast.
[Test of the concentration of purified plasmalogen derived from waste chicken peeled breast]
The concentration test was performed according to the method of Nadachi et al. ([Patent Document 1]). The concentration was 34% by mass.

Example 1
Conducting a clinical trial to examine the effects of cognitive function improvement by administration of a supplement containing an edible plasmalogen composition in carefully selected healthy Japanese subjects-a randomized, double-blind, placebo-controlled trial. It was outsourced to JACTA (Japan Clinical Trial Association Tokyo), which is a specialized agency in the field of clinical trials. The implementation period was 12 weeks from April 5th to June 29th, 2016, and was implemented in compliance with the ethical principles based on the Declaration of Helsinki. Consent forms for this study were obtained from all 135 healthy men and 135 healthy women aged 40 to 79 years.

I. Preparation of test sample 1) Preparation of test plasmalogen An edible plasmalogen composition in which 34% by mass PLs derived from peeled breast of abandoned chicken and an edible complex lipid are appropriately mixed according to Preparation Example 2-1. (Hereinafter referred to as "plasmalogen PLs-B").

2) Preparation of stock solutions for PLs 0.25 mg-containing soft capsules (hereinafter referred to as "plasmalogen EX") and PLs 0.50 mg-containing soft capsules (hereinafter referred to as "plasmalogen ES") by adding plasmalogen PLs-B. Plasmalogen EX and Plasmalogen ES stock solutions for soft capsules were prepared based on the formulations shown in Table 1.

3) Preparation of placebo stock solution for soft capsules Plasmalogen PLs-B was removed from the plasmalogen EX stock solution based on the formulation shown in Table 1, and starch was added to adjust the weight to prepare a placebo stock solution for soft capsules.
4) Soft encapsulation of each undiluted solution For the two types of test and the soft capsule for placebo, an ellipsoidal soft capsule coated with caramel-colored standard gelatin for food, which is indistinguishable from the appearance of each, was manufactured by an external specialized manufacturer. ..

II. Selection of subjects A total of 135 subjects were selected by document screening from the voluntary registrants of CROee Inc. (Tokyo) from March to April 2016.
1) Selection criteria;
(1) General healthy Japanese men and women aged 40 to 79 years (2) It is clear from the cognitive function interview shown in Fig. 1 that they have not taken appropriate medicines in the past even if they are not completely normal in terms of cognitive function. Who was made

2) Exclusion criteria; 54 excluded (1) Those who are being treated for cognitive dysfunction (2) Those who are taking drugs containing Chinese herbs (3) Pregnant women and women who may become pregnant during this study period ( 4) Those who are judged to be ineligible as subjects of this study by the person in charge of conducting this study

As a result of excluding 54 subjects from 135 subjects, the number of subjects was 81.
As a result, they were randomly assigned to three groups, group A (n = 26), group B (n = 28), and group C (n = 27). At this time, consideration was given so that the gender and age would not be biased.
* Group A is a placebo group * Group B is a test-1 (0.25 mg) group * Group C is a test-2 (0.5 mg) group The division is as above, and the subjects are 2 tablets daily (1 tablet after breakfast and dinner) (Each) soft capsules were ingested for 12 weeks while living a normal life, refraining from overdrinking and eating, and a diary describing the physical and forgetfulness was submitted to the supervisor of this study.

III. Test outline 1) Test schedule Table 2 shows the test schedule.

2) MMSE
See FIG. The highest score is 30.
3) Uchida-Krepperin test (hereinafter referred to as "UK test")
A simple single-digit addition is performed continuously for a certain period of time, and the calculation power test that determines "ability when a person calculates" and "characteristics when exerting that calculation power" based on the curve expressed by the amount of calculation. Is. Usually, it is performed in 2 sets of 1 minute x 15 times.
4) PSOL cognitive function self-diagnosis test (PSOL: represents P-solution)
See Figure 3 in a format that answers highly comprehensive questions about cognitive functionality that was uniquely devised. 2 is the reference point (baseline) on a 5-point scale from 0 to 4. Good evaluation when it exceeds 2.
5) Safety evaluation of the test sample (3 types of soft capsules) The evaluation was made by describing it in the daily report from the subject.

6) Data analysis method In this test, all analysis results were used without matching the number of samples.
All statistically processed values are expressed in mean ± SD.
Differences in measurements at weeks 0, 6 and 12 were statistically processed using the pair t-test. The comparative evaluation of the measured values of the MMSE, UK test, and PSOL cognitive function self-diagnosis test in the same group was performed using the pair t-test.

Student's t-test was used to make intergroup comparisons of measurements at weeks 0, 6 and 12 with deviations from baseline (weeks Δ0-6 and weeks Δ0-12). Background comparisons of the subjects in the group were performed using a one-way square deviation analysis.

We did not adjust for changes due to the timing of implementation. Subjects who mislisted were completely excluded from statistical processing. Statistical processing was performed using Statcel 4 and Excel statistics. The test statistical processing result between the two samples was judged to be significant at <5%.

IV. Test results 1) Statistical information of subjects 81 subjects were divided into 3 wards and the intake test was started, but 6 subjects dropped out. The reasons for this were 2 for poor physical condition, 3 for sudden work, and 1 for housework. In the end, 75 people completed the test. The breakdown was Test-1; 27, Test-2; 23, Placebo; 25, but there was no significant difference in age, gender and UK test (Table 3).

2) MMSE
The results are shown in FIG. 4 and Table 4. A significant difference was observed in the intra-group comparison of Test-1 and Test-2 at the 12th week. At the 6th week, a significant difference was observed in the comparison within the placebo group. However, no significant difference was observed between the test-1 vs. placebo and the test-2 vs. placebo in the group comparison.

3) UK test As shown in Fig. 5 and Table 4, no significant difference was observed in any of the three groups. However, in comparison between groups, a significant difference tended to be observed between Test-1 and placebo at 6 weeks.

4) The results of self-diagnosis of cognitive function are shown in Tables 5 to 8. In the comparison analysis of the amount of change between the groups, significant differences were found between the following items in Test-1 and Placebo, and in Test-2 and Placebo;
# 1 (Test-1, Test-2), # 5 (Test-1), # 7 (Test-1, Test-2), # 8 (Test-1), # 12 (Test-1) in the 6th week ), # 13 (Test-1), # 17 (Test-1, Test-2), # 18 (Test-1), # 25 (Test-1), # 26 (Test-2), and # 27 ( Test-1, Test-2),
On the other hand, in the 12th week, # 1 (Test-1), # 2 (Test-1, Test-2), # 4 (Test-1, Test-2), # 5 (Test-1, Test-2) , # 8 (Test-1, Test-2), # 12 (Test-1), # 15 (Test-2), # 16 (Test-2), # 17 (Test-2), # 20 (Test- 2), # 21 (Test-2), # 23 (Test-2), # 24 (Test-1, Test-2), # 26 (Test-2), and # 27 (Test-1, Test-2) ), A significant difference was observed in each of.
In Test-1 vs. Placebo, the number of significant difference items 10 in the 6th week decreased to 6 items in the 12th week, but in Test-2 vs. Placebo, the number of significant difference items 4 in the 6th week was significant in the 12th week. The number of difference items has increased significantly to 14.

5) Safety evaluation As far as the daily report is concerned, there is nothing that suggests doubts about the safety of the test sample, and there is no problem with safety.

V. Evaluation of test results 1) General remarks It was found that it is effective in improving cognitive function related to language and situation of healthy subjects who took a plasmalogen-containing supplement for 12 weeks. In addition, the test soft capsules did not interfere with safety during the 12-week test period.

There was no significant difference in the results of the UK test, but there was a significant difference between the high-dose intake group and the placebo group at 6 weeks.
Results suggest a dose dependence between the high-dose and low-dose groups, and a significant trend between them and the placebo group.
The dose is extremely low for foods and supplements, and it is on the order of "micron", which is considered to be less than oral medicine. In addition to being 0.5 mg or less per tablet, it is intended for healthy subjects and the administration period is 3 months. It is a surprising finding that the results suggesting an immediate improvement in computing power in a short time and in the 6th week.

2) Details (1) MMSE
A significant difference was observed within the 0.5 mg group and the 0.25 mg group at 12 weeks in each group. However, a significant difference was found within the placebo group at 6 weeks.
(2) UK test Immediately, a significant difference was observed between the high-dose group and the placebo group at 6 weeks, and a dose-dependent tendency was observed between the high-dose group and the low-dose group. Notable.
(3) PSOL cognitive function self-diagnosis test From the 12th week and the 6th week, significant differences were observed within and between the high and low dose groups and between the placebo groups. Although there were significant differences between the high and low dose groups, the results lacked dose dependence.

3) Overall evaluation * Random, double-blind, placebo-controlled trial
* Using an innovative edible plasmalogen composition,
* For carefully selected healthy subjects (135 document selections of institutional registrants ⇒ 81 cognitive function 3-level interview selections → 75 interview selections of examiners ⇒ 71 ineligible persons discovered during the examination)
* Extremely low daily doses, 0.5 mg and 1 mg, and
* Within a very short period of 12 weeks
A significant and immediate improvement effect on cognitive function was confirmed in just 6 weeks, and it was evaluated as a surprising result.

The fairness of the judgment described above is the title of IMPROVEMENT IN COGNITIVE FUNCTION BY SUPPLEMENT CONTAINED PLASMALOGEN FOR HEALTHY JAPANESE-A RAMDOMIZED, DOUBLE-BLIDED, PLASEBO-CONTROLED STUDY-, and is peer-reviewed "Clinical and New Drugs" Vol. 53, December It is published in the issue.

As described in detail above, the present invention relates to a safe and stable plasmalogen composition and a food for improving cognitive function (food composition) containing the same, and according to the present invention, 1) a safe organism. Provided is a safe and stable plasmalogen derived from chicken as a system material, and more specifically, an edible plasmalogen composition obtained by appropriately blending the plasmalogen and a purified complex lipid derived from chicken as an essential requirement. ) The effectiveness of a food for improving cognitive function (food composition) containing the plasmalogen composition for healthy subjects has been proved in a double-blind clinical study, and it is effective in delaying the onset of dementia in healthy subjects. To provide a food composition for safe, secure, inexpensive and convenient improvement of cognitive function, that is, specifically, to prevent forgetfulness related to the language and situation of healthy subjects, 3) Dementia Reserve Army It can be used industrially in that it alleviates the cognitive impairment, effectively suppresses the aggravation of the cognitive impairment, and contributes to the efficient reduction of the number of dementia patients. Has sex.

Claims (1)

Peeled breast of spawning abandoned chickens by forced molting (hereinafter referred to as "chicken peeled breast") is treated by an ethanol extraction method to contain at least diacylglycerophospholipid and sphingomyelin and dried. A complex lipid containing 10% by mass or more of ethanolamine-type plasmalogen (described as [PL-PE]) and choline-type plasmalogen (described as [PL-PC]) in terms of mass (hereinafter, “edible”). "Composite lipid") is prepared and
The edible complex lipid is enzymatically treated with phosphorlipase A1 (PLA1) to decompose diacylglycerophospholipids into free fatty acids and lysophospholipids, and these are removed to remove choline type [PL-PC]> ethanolamine type [PL-PE]. An edible plasmalogen having an ethanolamine-type and choline-type plasmalogen content of 20% by mass to 50% by mass in terms of dry mass, which is the plasmalogen composition derived from the peeled breast of chicken (hereinafter, "PLs"). ”) And prepare
The PLs and the edible complex lipid are appropriately mixed to prepare an edible plasmalogen composition containing both the PLs and the edible complex lipid (hereinafter, referred to as "edible PLs composition").
A stock solution for soft capsules or foodstuffs (hereinafter referred to as "edible PLs composition-δ-tocopherol stock solution") in which the edible PLs composition is uniformly mixed in δ-tocopherol having a mass of 100 to 200 times the mass thereof is used. Formulate,
It was related to the language and situation of a healthy subject containing the edible PLs composition containing the edible PLs composition-δ-tocopherol stock solution and δ-tocopherol containing 100 to 200 mass times the mass thereof as a base. A food composition to help you remember.