JP5496163B2 - Composition having an effect of preventing or ameliorating symptoms or diseases caused by a decrease in brain function - Google Patents

Description

  The present invention relates to a composition having an effect of preventing or ameliorating symptoms or diseases caused by a decrease in brain function, comprising arachidonic acid and / or a compound containing arachidonic acid as a constituent fatty acid as an active ingredient, and a method for producing the same. . More specifically, memory / learning comprising as an active ingredient at least one selected from the group consisting of arachidonic acid, alcohol esters of arachidonic acid, and triglycerides, phospholipids and glycolipids in which some or all of the constituent fatty acids are arachidonic acid Preventive or ameliorating agent for reduced ability, cognitive ability, emotional disorder (for example, depression), intellectual disorder (for example, dementia, specifically Alzheimer-type dementia, cerebrovascular dementia), and further preventive or ameliorating action And a method for producing the same.

  In recent years, with the advance of medical care, we are heading toward a rapidly aging society. Along with this, the number of senile dementia has also increased. According to the “2000 Health and Welfare White Paper” and “Dementia Elderly Countermeasure Review Report”, the number of demented elderly in 2000 was 1.5 to 1.6 million, and the number of demented patients aged 65 and over reached 1 in 14 ing. And by 2030, the number of patients is expected to increase steadily to 1 in 10 people. As senile dementia progresses, intellectual functions and emotions are impaired, resulting in problems in daily life and social life. By cause of senile dementia, it is divided into cerebrovascular dementia, Alzheimer type dementia, mixed type of them, research and development of drugs (cerebral circulation / metabolism improving drugs, anti-dementia drugs) effective in treating these brain diseases Unfortunately, there are currently no effective treatments available.

  As a person grows older, cerebral infarction develops to some extent, but the onset of dementia can be prevented, for example, by using the head. From this, it is considered possible to develop drugs aimed at prevention as well as treatment. However, drugs that can be taken easily from infants to the elderly, are safe, suppress brain function decline, prevent symptoms or diseases caused by brain function decline, and have further improved effects. Little developed.

  Traditionally, methods for improving brain function that have been studied are methods of improving the metabolism of brain energy that causes brain cells to efficiently absorb nutrients and activate the function of the cells (for example, increasing glucose in the brain). To improve cerebral circulation, improve cerebral circulation to supply brain cells with sufficient nutrients and oxygen (for example, increase in cerebral blood flow), and further through synaptic gaps via neurotransmitters Activation of neurotransmission (supplementation of precursors of neurotransmitters (eg, supplementation with choline, acetyl-CoA), inhibition of conversion of released neurotransmitters (eg, inhibition of acetylcholinesterase), neurotransmission Increased substance release (eg increased release of acetylcholine, glutamate, etc.), activation of neurotransmitter receptors, etc., and protection of neuronal membranes (eg antioxidants, membrane components) Replenishing, etc. prevention of arteriosclerosis) has been studied.

  In this way, in past studies, ingredients that prevent symptoms or diseases caused by decreased brain function and have further improvement effects have been found, but at present the effects are doubtful and effective drugs are also found in pharmaceuticals Not. Moreover, when considering application to foods, it has been difficult to be limited to ingredients derived from natural products.

  The brain is a tissue like a mass of lipids. For example, 1/3 in white matter and 1/4 in gray matter are occupied by phospholipids. The polyunsaturated fatty acids in phospholipids constituting various cell membranes of brain cells are mainly arachidonic acid and docosahexaenoic acid. However, these arachidonic acid and docosahexaenoic acid cannot be synthesized de novo in the animal body, and must be taken directly or indirectly (arachidonic acid, linoleic acid that is a precursor of docosahexaenoic acid, α-linolenic acid) from the diet. .

Therefore, attention has been focused on improving learning and memory ability of docosahexaenoic acid, prevention and recovery of senile dementia. However, not only docosahexaenoic acid, but also arachidonic acid is an important fatty acid that occupies a similar content in brain phospholipids. Sonderdegr et al. Show that the proportion of arachidonic acid in hippocampal phospholipids is significantly reduced to 8.1% by mass in Alzheimer patients compared to 12.4% by mass in normal individuals (Lipids 26 , 421-425 ( 1991)). Thus, although arachidonic acid suggests the possibility of playing an important role in maintaining brain function, there has been no specific demonstration since there was not a sufficient source of arachidonic acid.

  Several inventions that use arachidonic acid to maintain brain function have been shown. "Learning ability improver" described in JP-A-6-256179 is an invention comprising a 1,2-diacyl-sn-glycerol derivative as an active ingredient, and various polyunsaturated fatty acids bonded to the 2-position are listed. As one of them, arachidonic acid is shown. However, only the 1,2-diacyl-sn-glycerol derivatives bound with docosahexaenoic acid are specifically shown in the examples, and arachidonic acid is merely listed, and the effect is completely It has not been demonstrated.

  In the “brain function improving and nutritional composition” described in JP-A-10-101568, a combination of ganglioside and arachidonic acid is shown as a means for providing a novel brain function improving agent and a nutritional composition containing the same. Yes. However, although experiments with spontaneously aged rats are shown as test examples, the age of the test is only 13 months old, equivalent to 33 years for humans (one day for a rat corresponds to one month for humans) This is not an aging model test. The ratio of arachidonic acid or the amount of arachidonic acid in brain phospholipids generally does not change at this age, and arachidone does not show a decrease in brain function due to aging. It is common to think that acid effects cannot occur. Actually, the effects of arachidonic acid alone have not been evaluated in the test examples, and only show that arachidonic acid enhances the effect of ganglioside.

  “Protein kinase C isozyme activator” described in JP-A-6-279511 activates protein kinase C, which plays an important role in intracellular signal transduction, and has a therapeutic agent for senile dementia as an accompanying effect. Show. However, the active ingredient is a phosphatidylserine derivative having a highly unsaturated fatty acid as a constituent fatty acid, and one of the highly unsaturated fatty acids is arachidonic acid. However, in the examples, the effect of arachidonic acid is not much different from that of phosphatidylserine derivatives bound with linoleic acid and α-linolenic acid, and there is no advantage of phosphatidylserine derivatives having arachidonic acid as a constituent fatty acid. Has not been demonstrated.

  Further, the evaluation is based merely on the measurement of enzyme activity, and no effect of preventing or ameliorating symptoms or diseases caused by a decrease in brain function has been clarified. As described above, although several inventions using arachidonic acid for maintaining the function of the brain have been shown, animal experiments were conducted because arachidonic acid and a compound containing arachidonic acid as a constituent fatty acid did not exist in a sufficient supply amount. Thus, the true effect could not be clarified, and arachidonic acid was merely described as one of the fatty acids, and the actual situation was not a true invention.

  A pathological memory disorder associated with organic lesions in the brain. Short- and long-term memory impairment is a core symptom of dementia. However, forgetfulness, which expresses memory impairment in other words, is one of the most common complaints in the elderly, and various studies have pointed out that human learning and memory decline with physiological aging. (Katzman, R. and Terry, R., The Neurology of Aging, FA Davis, Philadelphia, pp. 15-50).

  When the memory is viewed from the time course that it is formed, it is classified into sensory memory, primary memory, and secondary memory. Primary memory is sometimes called immediate memory, and secondary memory is sometimes called long-term memory. Short-term memory may refer to primary memory or to learning ability across secondary memory. Sensory memory is formed when there is visual input lasting about 50 milliseconds, but it is extremely unstable and disappears within only 250-500 milliseconds. The primary memory is always kept conscious while information is cognitively processed, and has a role as a working memory.

  Information that has undergone processing as primary storage is transferred to secondary storage and held for a long period of time, and then recollected through primary storage. The capacity of primary memory is small and will disappear in 20-30 seconds unless recollected repeatedly. Secondary storage consists of the steps of transferring and consolidation of information processed in the primary storage, its semi-permanent storage, and its retrieval. This secondary memory is severely impaired with aging. This decrease in secondary memory is mainly due to a failure in the stage until storage of the memory, and there is almost no decline in the ability to reproduce the memory stored from a young age. However, dementia patients always suffer from this memory.

  One effect of arachidonic acid has been clarified as an electrophysiological analysis. It is known that synapses are excited when high-frequency stimulation is applied to the brain hippocampus, and the subsequent synaptic response is high. This phenomenon is called hippocampal LTP (long-term potentiation), and is a phenomenon that is the basis of synaptic plasticity, and is an index of brain function evaluation. BM McGahon et al. Measured LTP in the rat hippocampus in 22-month-old aged rats fed with a control diet or arachidonic acid-containing diet (10 mg / rat / day) for 8 weeks (Neurobiol Aging 20, 643 (1999) ). The hippocampal LTP in the old rat is clearly reduced compared to the 4-month-old young rat, indicating that arachidonic acid administration restores to the level of the young rat.

  However, the enhancement of hippocampal LTP indicates the activation of primary memory in terms of the memory mechanism, not the activation of the transition from primary memory to secondary memory necessary for memory fixation. Thus, memory consolidation cannot be demonstrated unless revealed by behavioral pharmacology studies. Although an example of evaluating the effect of arachidonic acid with an electrophysiological index is shown in this way, arachidonic acid and / or arachidonic acid is used for the prevention or improvement of symptoms or diseases caused by the decrease in brain function of the present invention. It has not been clarified whether the compound as a constituent fatty acid is effective.

  Accordingly, there is a strong demand for the development of safer compounds that prevent symptoms or diseases caused by a decrease in brain function, exhibit further improvement effects, and are excellent in application to pharmaceuticals and foods.

  Accordingly, the present invention provides an agent for preventing or ameliorating a symptom or disease caused by a decrease in brain function, a symptom resulting from a decrease in brain function, or an arachidonic acid and / or a compound containing arachidonic acid as a constituent fatty acid. An object of the present invention is to provide a food and drink having a disease prevention or ameliorating action and a method for producing the same. More specifically, memory / learning comprising as an active ingredient at least one selected from the group consisting of arachidonic acid, alcohol esters of arachidonic acid, and triglycerides, phospholipids and glycolipids in which some or all of the constituent fatty acids are arachidonic acid Preventive or ameliorating agent for reduced ability, cognitive ability, emotional disorder (for example, depression), intellectual disorder (for example, dementia, specifically Alzheimer-type dementia, cerebrovascular dementia), and further preventive or ameliorating action It is going to provide the food-drinks which have, and its manufacturing method.

  As a result of intensive research for the purpose of clarifying the preventive or ameliorating effect on symptoms or diseases caused by a decrease in brain function of arachidonic acid or a compound containing arachidonic acid as a constituent fatty acid, the present inventors have surprisingly found that Old rats over 20 months old were subjected to a Morris water maze test, and the effects of arachidonic acid or a compound containing arachidonic acid as a constituent fatty acid were clarified by behavioral pharmacological analysis.

Furthermore, the present inventors have succeeded in industrial production of triglycerides containing arachidonic acid in an amount of 20% by mass or more by microorganisms, which can be used for the effect test of the present invention, and revealed the effects of the triglycerides.
Furthermore, the present inventors have succeeded in producing an oil and fat containing a triglyceride in which a medium chain fatty acid is bonded to the 1,3-position and arachidonic acid is bonded to the 2-position by an enzymatic method, and used for the effect test of the present invention. And the effect of the triglyceride was clarified.

  Accordingly, the present invention provides an agent for preventing or ameliorating a symptom or disease caused by a decrease in brain function, a symptom resulting from a decrease in brain function, or an arachidonic acid and / or a compound containing arachidonic acid as a constituent fatty acid. Provided are foods and drinks having a disease-preventing or ameliorating action and methods for producing the same. More specifically, memory / learning comprising as an active ingredient at least one selected from the group consisting of arachidonic acid, alcohol esters of arachidonic acid, and triglycerides, phospholipids and glycolipids in which some or all of the constituent fatty acids are arachidonic acid Preventive or ameliorating agent for reduced ability, cognitive ability, emotional disorder (for example, depression), intellectual disorder (for example, dementia, specifically Alzheimer-type dementia, cerebrovascular dementia), and further preventive or ameliorating action The food and drink which have this, and its manufacturing method are provided.

FIG. 1 is a schematic explanatory diagram of an apparatus used for a Morris water maze test. FIG. 2 illustrates learning acquisition (Hit%). FIG. 3 is a graph showing learning acquisition (Hit%) with respect to the number of trials of the rat. FIG. 4 is a diagram showing a trajectory for 60 seconds when a rat swims in a probe test for measuring the degree of acquisition of learning.

FIG. 5 is a graph showing the results of a probe test for measuring the degree of acquisition of learning. FIG. 6 is a graph showing the results of the correlation between learning parameters and the amount of arachidonic acid in the hippocampus.

  The present invention relates to a preventive or ameliorating agent for a symptom or disease caused by a decrease in brain function, and a symptom or disease caused by a decrease in brain function, comprising arachidonic acid and / or a compound containing arachidonic acid as a constituent fatty acid as an active ingredient. It is related with the food-drinks which have the prevention or improvement effect of this, and its manufacturing method.

  Since the composition of the present invention has a preventive or ameliorating effect on symptoms or diseases caused by a decrease in brain function, for example, a decrease in memory / learning ability, a decline in cognitive ability, an emotional disorder (for example, depression etc.), It is useful as a food / drink, pharmaceutical, quasi-drug, etc. for the purpose of prevention / improvement (or treatment) of intellectual disability (for example, dementia, specifically Alzheimer-type dementia, cerebrovascular dementia) .

  More specifically, the composition of the present invention has an effect of preventing or ameliorating symptoms or diseases caused by a decrease in brain function associated with aging, a decrease in memory / learning ability, a decline in cognitive ability, emotions Food and drinks, pharmaceuticals for the purpose of prevention / improvement (or treatment) of disorders (eg depression), intellectual disorders (eg dementia, specifically Alzheimer-type dementia, cerebrovascular dementia), Quasi-drugs, as well as prevention of forgetfulness, prevention of blurring, maintenance / improvement of memory, maintenance / improvement of concentration, maintenance / improvement of attention, refreshment of head, renewal of head, rejuvenation, etc. It is useful as foods and drinks, health foods, functional foods, foods for specified health use, foods for the elderly, etc.

  As the active ingredient of the present invention, in addition to free arachidonic acid, all compounds containing arachidonic acid as a constituent fatty acid can be used. Examples of the compound having arachidonic acid as a constituent fatty acid include arachidonic acid salts such as calcium salts and sodium salts. Further, alcohol esters of arachidonic acid such as arachidonic acid methyl ester and arachidonic acid ethyl ester can be used. In addition, triglycerides, phospholipids, and glycolipids in which some or all of the constituent fatty acids are arachidonic acid can be used.

  When considering application to foods, it is desirable that arachidonic acid be in the form of triglycerides or phospholipids, particularly triglycerides. Although there was almost no natural source of triglycerides containing arachidonic acid (synonymous with triglycerides containing triglycerides in which some or all of the constituent fatty acids are arachidonic acid), the inventors have included arachidonic acid The effect of the active ingredient of the present invention was revealed for the first time by behavioral pharmacological analysis by subjecting an aged rat over 20 months old to the Morris water maze test. Clarified that it has a preventive or ameliorating effect on symptoms or diseases caused by reduced function.

  Therefore, in this invention, the triglyceride (triglyceride containing arachidonic acid) containing the triglyceride whose one part or all part of the constituent fatty acid which is an active ingredient of this invention is arachidonic acid can be used. The triglyceride containing arachidonic acid is an oil or fat having a ratio of arachidonic acid of 20% by mass or more, preferably 30% by mass or more, more preferably 40% by mass or more of all fatty acids constituting the triglyceride ( Triglyceride) is a desirable form when applied to food. Therefore, in the present invention, any microorganisms obtained by culturing microorganisms having the ability to produce oils and fats (triglycerides) containing arachidonic acid can be used.

The microorganism capable of producing fat or oil (triglyceride) containing arachidonic acid, Mortierella (Mortierella) genus, Koni audio bolus (Conidiobolus) genus Fichiumu (Pythium) genus Phytophthora (Phytophthora) genus Penishiryumu (Penicillium) genus , Clos de sports Liu beam (Cladosporium) genus, Mucor (Mucor) genus, Fuzaryumu (Fusarium) genus Aspergillus (Aspergillus) genus, Rhodotorula (Rhodotorula) genus, Entomofutora (Entomophthora) genus, echinochrome aminocephalosporanic indium (Echinosporangium) genus Saprolegnia (Saprolegnia And) microorganisms belonging to the genus.

In the Mortierella (Mortierella) belonging to the genus Mortierella (Mortierella) microorganisms belonging to the subgenus, for example Mortierella elongata (Mortierella elongata), Mortierella exigua (Mortierella exigua), Mortierella Figurofira (Mortierella hygrophila), Mortierella alpina ( Mortierella alpina ). More specifically, Mortierella elongata IFO8570, Mortierella exigua IFO8571, Mortierella hygrophila IFO5941, Mortierella alpina 221266266, CC41, 266266CC Examples include CBS219.35, CBS224.37, CBS250.53, CBS343.66, CBS527.72, CBS529.72, CBS608.70, CBS754.68 and the like.

  All of these strains are obtained from Osaka City Foundation for Fermentation (IFO), American Type Culture Collection (ATCC), and Centrralbureau voor Schimmelcultures (CBS) without any restrictions. can do. In addition, the strain Mortierella elongata SAM0219 (Mikken Kenyoku No. 8703) (Mikken Kenjyo No. 1239) isolated from soil by the research group of the present invention can also be used.

  In order to culture the strain used in the present invention, the spore, mycelium of the strain or a preculture obtained by culturing in advance is inoculated into a liquid medium or a solid medium and cultured. In the case of a liquid medium, commonly used carbon sources such as glucose, fructose, xylose, saccharose, maltose, soluble starch, molasses, glycerol, mannitol, etc. can be used, but are not limited to these. It is not a thing.

  Nitrogen sources include natural nitrogen sources such as peptone, yeast extract, malt extract, meat extract, casamino acid, corn steep liquor, soy protein, defatted soybean, cottonseed residue, etc., organic nitrogen sources such as urea, and sodium nitrate Inorganic nitrogen sources such as ammonium nitrate and ammonium sulfate can be used. In addition, inorganic salts such as phosphate, magnesium sulfate, iron sulfate, and copper sulfate, vitamins, and the like can be used as a trace nutrient source as necessary. These medium components are not particularly limited as long as they do not impair the growth of microorganisms. Generally, in practice, the carbon source has a concentration of 0.1 to 40 mass (W / V)%, preferably 1 to 25 mass (W / V)%. The initial nitrogen source addition amount is 0.1 to 10 mass (W / V)%, preferably 0.1 to 6 mass (W / V)%, and the nitrogen source may be fed during the cultivation.

  Furthermore, by controlling the medium carbon source concentration, an oil (triglyceride) containing 45% by mass or more of arachidonic acid can be used as the active ingredient of the present invention. In the culture, the cell growth phase is from the 2nd to 4th day of the culture, and the oil accumulation phase is from the 2nd to 4th day of the culture. The initial carbon source concentration is 1 to 8% by mass, preferably 1 to 4% by mass, and the carbon source is added sequentially only during the initial period of the cell growth phase and the fat accumulation phase, and the total of the carbon sources added sequentially is added. Is 2 to 20% by mass, preferably 5 to 15% by mass. In addition, the sequential addition amount of the carbon source during the cell growth phase and the early stage of the fat accumulation phase is added according to the initial nitrogen source concentration, more preferably after the 7th day of culture, preferably after the 6th day of culture. Makes it possible to obtain an oil (triglyceride) containing 45% by mass or more of arachidonic acid by setting the carbon source concentration in the medium after the fourth day of culture to 0, and can be used as an active ingredient of the present invention. it can.

  The culture temperature of the arachidonic acid-producing bacterium varies depending on the microorganism to be used, but it is 5 to 40 ° C., preferably 20 to 30 ° C. After culturing at 20 to 30 ° C. to grow the cells, 5 to 20 ° C. The unsaturated fatty acid can also be produced by continuing the culture at Such a temperature control can also increase the proportion of highly unsaturated fatty acids in the produced fatty acids. The pH of the medium is 4 to 10, preferably 5 to 9, and aeration stirring culture, shaking culture, or stationary culture is performed. The culture is usually performed for 2 to 30 days, preferably 5 to 20 days, more preferably 5 to 15 days.

Furthermore, as a means to increase the ratio of arachidonic acid in fats and oils (triglycerides) containing arachidonic acid, in addition to controlling the medium carbon source concentration, the arachidonic acid-containing fats and oils are selectively hydrolyzed to obtain high arachidonic acid-containing fats and oils. It can also be obtained. The lipase used for this selective hydrolysis does not have triglyceride regiospecificity, and its hydrolysis activity decreases in proportion to the number of double bonds, so that ester bonds of fatty acids other than highly unsaturated fatty acids are hydrolyzed. . Then, transesterification occurs between the produced PUFA partial glycerides, resulting in triglycerides with highly unsaturated fatty acids (“Enhancement of Archidonic: Selective Hydrolysis of a Single-Cell Oil from Mortierella with Candida cylindracea Lipase” : J. Am. Oil Chem. Soc., 72 , 1323-1327 (1998)).

  Thus, fats and oils (triglycerides) containing a high amount of arachidonic acid obtained by selective hydrolysis of arachidonic acid-containing fats and oils can be used as the active ingredient of the present invention. The ratio of arachidonic acid to the total fatty acids of the oil (triglyceride) containing arachidonic acid of the present invention is preferably high for the purpose of eliminating the influence of other fatty acids, but is not limited to a high ratio, When applied to foods, the absolute amount of arachidonic acid may be a problem, and even oils (triglycerides) containing 10% by mass or more of arachidonic acid can be used substantially.

  Furthermore, in the present invention, as a triglyceride in which some or all of the constituent fatty acids are arachidonic acid, a triglyceride in which a medium chain fatty acid is bonded to the 1,3-position and arachidonic acid is bonded to the 2-position can be used. Further, a medium chain fatty acid in the 1,3-position and a triglyceride in which arachidonic acid is bonded in the 2-position are 5 mol% or more, preferably 10 mol% or more, more preferably 20 mol% or more, most preferably 30 mol%. Oils and fats (triglycerides) containing the above can be used. The medium chain fatty acid bonded to the 1,3-position of the triglyceride may be selected from fatty acids having 6 to 12 carbon atoms. Examples of the fatty acid having 6 to 12 carbon atoms include caprylic acid and capric acid, and 1,3-capryloyl-2-arachidonoyl-glycerol (hereinafter also referred to as “8A8”) is preferable.

  These triglycerides in which a medium chain fatty acid is bonded to the 1,3-position and arachidonic acid is bonded to the 2-position are optimal oils and fats (triglycerides) when targeting the elderly. In general, fats and oils (triglycerides) ingested are hydrolyzed by pancreatic lipase when entering the small intestine, but this pancreatic lipase is specific to the 1,3-position, and the 1,3-position of the triglyceride is cut off. A molecular free fatty acid is formed, and at the same time, one molecule of 2-monoacylglycerol (hereinafter referred to as “2-MG”) is produced. Since this 2-MG has very high bile acid solubility and good absorbability, it is generally said that the 2-position fatty acid has better absorbability.

  In addition, 2-MG acts as a surfactant when dissolved in bile acids, and acts to increase the absorbability of free fatty acids. Next, free fatty acids and 2-MG are biosynthesized with bile acid complex micelles together with cholesterol and phospholipids and taken into small intestinal epithelial cells, re-synthesis of triacylglycerol occurs, and finally it is lymphatic as chylomicron. Will be released. However, fatty acid characteristics of this pancreatic lipase are higher than saturated fatty acids, and arachidonic acid is difficult to cut. A further problem is that pancreatic lipase activity decreases with age, so that middle-chain fatty acids are located in the 1,3-position and 2- The triglyceride in which arachidonic acid is bound is an optimal oil (triglyceride).

  As one of the specific methods for producing triglycerides in which medium chain fatty acid is bonded to 1,3-position and arachidonic acid is bonded to 2-position, triglyceride in the presence of oil (triglyceride) containing arachidonic acid and medium chain fatty acid Can be produced by acting a lipase that specifically acts on the ester bond at the 1,3-position.

  Oils and fats (triglycerides) used as raw materials are triglycerides containing arachidonic acid as a constituent fatty acid. When the ratio of arachidonic acid to the total fatty acids constituting the triglycerides is high, unreacted fats and oils (raw triglycerides and fatty acids at 1,3-position) In order to prevent a decrease in the reaction yield due to an increase in triglycerides in which only one of them is a medium chain fatty acid), the enzyme reaction temperature is higher than the usual enzyme reaction temperature 20-30 ° C, preferably 30-50 ° C, preferably 40-50 ° C .

Examples of lipases that specifically act on the ester bond at the 1,3-position of triglycerides, such as those produced by microorganisms such as Rhizopus , Rhizomucor , Aspergillus , porcine pancreatic lipase, etc. Can be mentioned. A commercially available lipase can be used. For example, lipase of Rhizopus delemar (Rhizopus delemar) (Tanabe Seiyaku Co., Ltd., Talipase), Rhizomucor Mihai (Rhizomucor miehei) lipase (Novo Nordisk Co., Ltd., a ribozyme IM), Aseperugirusu niger (Apergillus niger ) lipase (manufactured by Amano Pharmaceutical Co., Ltd., lipase A) and the like, but are not limited to these enzymes, and any lipase specific to the 1,3-position can be used.

  In order to increase the reaction efficiency, the lipase is used at a temperature of 30 ° C. or higher, preferably 40 ° C. or higher. Is desirable. Examples of the immobilization carrier include a porous (hypoporous) resin and an ion exchange resin carrier having a pore diameter of about 100 angstroms or more, such as Dowex MARATHON WBA (trademark, Dow Chemical).

  The immobilized carrier 1 is suspended in an aqueous solution of 1,3-position specific lipase 0.5 to 20 times (mass), and 2 to 5 times the amount of cold acetone (for example, -80 ° C) is added to the suspension. Add slowly with stirring to form a precipitate. This precipitate can be dried under reduced pressure to prepare an immobilized enzyme. In a simpler method, 0.05-0.4 times the amount of 1,3-position-specific lipase is dissolved in a minimum amount of water with respect to the immobilized carrier 1, and the immobilized carrier is mixed with stirring, under reduced pressure. The immobilized enzyme can be prepared by drying. By this operation, about 90% of the lipase is immobilized on the carrier, but as it is, the transesterification activity is not shown at all, and a substrate (raw oil and medium chain fatty acid) added with 1 to 10% (w / v) water is added. Among them, the immobilized enzyme can be activated most efficiently by being pretreated in a substrate to which 1 to 3% by mass of water is added, and can be used for production.

  Depending on the type of enzyme, the amount of water added to the reaction system is extremely important. If water is not included, transesterification will not proceed easily, and if the amount of water is high, hydrolysis will occur and glyceride recovery will occur. (If hydrolysis occurs, diglycerides and monoglycerides are produced). However, in this case, by using the immobilized enzyme activated by the pretreatment, the amount of water added to the reaction system is not important, and a transesterification reaction can be efficiently performed even in a system containing no water at all. Further, the pretreatment can be omitted by selecting the kind of the enzyme agent.

  In this way, by using an immobilized enzyme with heat resistance and raising the enzyme reaction temperature, the reaction efficiency of fats and oils (triglycerides) containing arachidonic acid with low reactivity of the 1,3-position specific lipase is also achieved. Without lowering, it is possible to efficiently produce a triglyceride in which a medium chain fatty acid is bonded to the 1,3-position and arachidonic acid is bonded to the 2-position.

  In a method for producing a food or drink having an effect of preventing or ameliorating symptoms or diseases caused by a decrease in brain function, arachidonic acid and / or a compound containing arachidonic acid as a constituent fatty acid alone or substantially containing arachidonic acid Even if it is contained, it can be blended together with a small amount of raw material for food and drink. Here, even if arachidonic acid is contained in the raw material for food and drink, even if a person ingests the food composition containing the arachidonic acid, the daily intake of arachidonic acid according to the present invention described later is used. It means the amount that does not reach the amount.

  In particular, in the case of triglycerides in which some or all of the constituent fatty acids are arachidonic acid, there are endless possibilities for the use of oils and fats (triglycerides), and as raw materials and additives for foods, beverages, pharmaceuticals, quasi drugs It can be used. And there is no restriction on the purpose and amount of use.

  Examples of food compositions include functional foods, special health foods, nutritional supplements, premature infant formulas, infant formulas, infant foods, maternal foods or elderly foods in addition to general foods. be able to. Examples of foods that contain fats and oils include natural foods that originally contain fats and oils such as meat, fish, and nuts, foods that add fats and oils during cooking such as soups, foods that use fats and oils as a heating medium such as donuts, fats and foods such as butter, and cookies Processed foods to which fats and oils are added at the time of processing such as, or foods to which fats and oils are sprayed or applied at the time of processing and finishing of hard biscuits and the like. Furthermore, it can be added to agricultural foods, fermented foods, livestock foods, marine foods, or beverages that do not contain fats and oils. Furthermore, it may be in the form of functional foods, pharmaceuticals, quasi drugs, such as enteral nutrients, powders, granules, tablets, capsules, troches, liquids for internal use, suspensions, emulsions, syrups. It may be a processing form such as.

  In addition to the active ingredient of the present invention, the composition of the present invention may contain various carriers and additives generally used for foods and drinks, pharmaceuticals or quasi drugs. In particular, it is desirable to contain an antioxidant for the purpose of preventing oxidative degradation of the active ingredient of the present invention. Antioxidants include, for example, tocopherols, flavone derivatives, phyllozurcins, kojic acid, gallic acid derivatives, catechins, fuxic acid, gossypol, pyrazine derivatives, sesamol, guaiacol, guaiac fat, p-coumaric acid, nordihydroguayate Natural antioxidants such as tic acids, sterols, terpenes, nucleobases, carotenoids, lignans and ascorbyl palmitate, ascorbate stearate, butylhydroxyanisole (BHA), butylhydroxytoluene ( BHT), mono-t-butylhydroquinone (TBHQ), and 4-hydroxymethyl-2,6-di-t-butylphenol (HMBP).

  Examples of tocopherols include α-tocopherol, β-tocopherol, γ-tocopherol, δ-tocopherol, ε-tocopherol, ξ-tocopherol, η-tocopherol and tocopherol esters (such as tocopherol acetate). Further, examples of carotenoids include β-carotene, canthaxanthin, astaxanthin and the like.

  In addition to the active ingredient of the present invention, the composition of the present invention includes various carriers, extenders, diluents, extenders, dispersants, excipients, binder solvents (eg, water, ethanol, vegetable oil) as carriers. ), Solubilizers, buffers, solubilizers, gelling agents, suspending agents, wheat flour, rice flour, starch, corn starch, polysaccharides, milk protein, collagen, rice oil, lecithin and the like. Examples of additives include vitamins, sweeteners, organic acids, colorants, flavors, anti-wetting agents, fibers, electrolytes, minerals, nutrients, antioxidants, preservatives, fragrances, wetting agents, natural foods An extract, a vegetable extract, etc. can be mentioned, but it is not necessarily limited to these.

  Arachidonic acid is the main medicinal component of arachidonic acid and compounds containing arachidonic acid as a constituent fatty acid. The daily intake of arachidonic acid from meals was reported to be 0.14 g in the Kanto area and 0.19 to 0.20 g in the Kansai area (Lipid Nutrition 4, 73-82, 1995). It is necessary to ingest a considerable amount or more of arachidonic acid from the viewpoints of lowering the lipase activity and pancreatic lipase activity. Therefore, the daily intake of an arachidonic acid and a compound containing arachidonic acid as a constituent fatty acid of the present invention per day (for example, assuming a body weight of 60 kg) is 0.001 g to 20 g, preferably 0.01 g to 10 g, in terms of arachidonic acid. More preferably 0.05 to 5 g, and most preferably 0.1 to 2 g.

  When the active ingredient of the present invention is actually applied to a food or drink, the absolute amount of arachidonic acid to be blended in the food is also important. However, since the absolute amount to be blended in food and drink also varies depending on the amount of food and drink to be blended, when blending triglycerides containing triglycerides in which some or all of the constituent fatty acids are arachidonic acid, Arachidonic acid is blended so as to be 0.0003% by mass or more, preferably 0.003% by mass or more, more preferably 0.03% by mass or more. Furthermore, when a triglyceride containing a triglyceride in which a medium chain fatty acid is bonded to the 1,3-position and an arachidonic acid is bonded to the 2-position, the medium chain fatty acid is added to the 1,3-position, The triglyceride having arachidonic acid bonded to the position is added in an amount of 0.001% by mass or more, preferably 0.01% by mass or more, more preferably 0.1% by mass or more.

When the composition of the present invention is used as a pharmaceutical, it can be produced according to a method commonly used in the pharmaceutical technical field, for example, a method described in the Japanese Pharmacopoeia or a method analogous thereto.
When using the composition of this invention as a pharmaceutical, the compounding quantity of the active ingredient in a composition is not specifically limited as long as the objective of this invention is achieved, It can be used in a suitable mixture ratio suitably.
When the composition of the present invention is used as a pharmaceutical, it is desirable to administer it in a dosage unit form, and oral administration is particularly preferred. The dose of the composition of the present invention varies depending on age, body weight, symptoms, frequency of administration, etc., for example, an adult (about 60 kg) per day, arachidonic acid of the present invention and a compound containing arachidonic acid as a constituent fatty acid, In terms of the amount of arachidonic acid, usually about 0.001 g to 20 g, preferably about 0.01 g to 10 g, more preferably about 0.05 to 5 g, and most preferably about 0.1 g to 2 g are administered in 1 to 3 divided doses per day. It is good.

  The main fatty acids of the brain phospholipid membrane are arachidonic acid as well as docosahexaenoic acid, and considering the balance, the composition of the present invention desirably combines arachidonic acid with docosahexaenoic acid. Moreover, since the ratio of eicosapentaenoic acid is very low in the phospholipid membrane of the brain, it is desirable that the composition of the present invention hardly contains eicosapentaenoic acid. A composition containing almost no eicosapentaenoic acid and containing arachidonic acid and docosahexaenoic acid is more desirable. In the combination of arachidonic acid and docosahexaenoic acid, the arachidonic acid / docosahexaenoic acid ratio (mass) is preferably in the range of 0.1 to 15, and preferably in the range of 0.25 to 10. Moreover, the food / beverage which mix | blended the amount of eicosapentaenoic acid of the quantity which does not exceed 1/5 (mass ratio) of arachidonic acid is desirable.

Next, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to the following examples.
In addition, in the food composition of the present invention such as food and drink, health food, functional food, food for specified health use, food for the aged, etc., the food composition and / or a component in the food composition is reduced in brain function. Preventive or ameliorating symptoms or diseases caused by symptoms, more specifically, preventing or ameliorating symptoms or diseases caused by aging-related decline in brain function, decline in memory / learning ability, decline in cognitive ability, emotion Prevention / improvement of disability (eg, depression), intellectual disability (eg, dementia, specifically Alzheimer-type dementia, cerebrovascular dementia), prevention of forgetfulness, prevention of blurring, maintenance / improvement of memory, concentration Selling the food composition and / or the food composition to maintain / improve power, maintain / improve attention, clean the head, frighten the head, rejuvenate, etc. Tools to promote For example also include those sold by, for example, described or displayed in brochures).

Example 1. Method for Producing Triglyceride Containing Arachidonic Acid Mortierella alpina CBS754.68 was used as an arachidonic acid-producing bacterium. 1.8% glucose, defatted soybean flour 3.1%, 0.1% soybean _{oil, KH 2 PO 4 0.3%,} Na 2 SO 4 0.1%, medium with CaCl ₂ · 2H ₂ O 0.05% and _{MgCl 2 · 6H 2 O 0.05%} 6kL Was prepared in a 10 kL culture tank, and the initial pH was adjusted to 6.0. 30 L of the preculture was inoculated and aerated and stirred for 8 days under conditions of a temperature of 26 ° C., an aeration rate of 360 m ³ / hour, and a tank pressure of 200 kPa. The stirring number was adjusted so that the dissolved oxygen concentration was maintained at 10 to 15 ppm.

  Further, the glucose concentration was maintained at 0.5 to 1% until the fourth day so that the glucose concentration in the medium was within the range of 1 to 2.5% by fed-batch method (the above% is the mass (w / v)%)). After completion of the culture, the cells containing triglycerides containing arachidonic acid are collected by filtration and drying, and the fats and oils are extracted from the obtained cells with hexane, and the edible fats and oils purification process (degumming, deoxidation, deodorization, After decolorization, 150 kg of arachidonic acid-containing triglyceride (arachidonic acid bound to an arbitrary position of the triglyceride) was obtained.

  The obtained oil (triglyceride) was methyl esterified, and the obtained fatty acid methyl ester was analyzed by gas chromatography. As a result, the ratio of arachidonic acid in the total fatty acid was 40.84% by mass. Palmitic acid, stearic acid, oleic acid, linoleic acid, γ-linolenic acid, dihomo-γ-linolenic acid and the like were 11.63, 7.45, 7.73, 9.14, 2.23, and 3.27% by mass, respectively. Further, the above arachidonic acid-containing oil (triglyceride) was ethyl esterified, and 99% by mass of arachidonic acid ethyl ester was separated and purified from a fatty acid ethyl ester mixture containing 40% by mass of arachidonic acid ethyl ester by a conventional high performance liquid chromatography. .

Example 2. Production of triglyceride containing 5A mol or more of 8A8 100 g of ion exchange resin carrier (Dowex MARATHON WBA: Dow Chemical, Trademark) in 80 ml of Rhizopus delemar lipase 12.5% aqueous solution (Tallipase powder: Tanabe Seiyaku Co., Ltd.) The suspension was suspended and dried under reduced pressure to obtain an immobilized lipase.

  Next, 80 g of triglyceride (TGA40S) containing 40% by mass of arachidonic acid obtained in Example 1, 160 g of caprylic acid, 12 g of the above immobilized lipase, and 4.8 ml of water were reacted at 30 ° C. for 48 hours with stirring (130 rpm). I let you. After completion of the reaction, the reaction solution was removed to obtain an activated immobilized lipase.

Next, 10 g of immobilized lipase ( Rhizopus delemar lipase, carrier: Dowex MARATHON WBA ™) was packed in a jacketed glass column (1.8 x 12.5 cm, capacity 31.8 ml), and TGA40S obtained in Example 1 and caprylic acid were loaded. The mixed fat / oil mixed 1: 2 was allowed to flow through the column at a constant flow rate (4 ml / h), and a continuous reaction was carried out to obtain 400 g of the reactive fat / oil. The column temperature was 40 to 41 ° C. Unreacted caprylic acid and free fatty acids were removed from the obtained reaction fats and oils by molecular distillation to obtain fats and oils (triglycerides) containing 8A8.

  And when the ratio of 8A8 in the obtained 8A8-containing oil (triglyceride) was examined by gas chromatography and high performance liquid chromatography, it was 31.6 mol% (Note that 8P8, 808, 8L8, 8G8, 8D8 The ratios were 0.6, 7.9, 15.1, 5.2, and 4.8 mol%, respectively, and fatty acids P, O, L, G, and D that bind to the 2-position of triglyceride were palmitic acid, oleic acid, linoleic acid, and γ-linolene, respectively. Acid, dihomo-γ-linolenic acid, 8P8 is 1,3-capryloyl-2-palmitoyl-glycerol, 808 is 1,3-capryloyl-2-oleoyl-glycerol, 8L8 is 1,3-capryloyl-2- Linoleoyl-glycerol, 8G8 refers to 1,3-capryloyl-2-γ-linolenoyl-glycerol, and 8D8 refers to 1,3-caprylyl-2-dihomo-γ-linolenoyl-glycerol. In addition, 96 mol% 8A8 was separated and purified from the obtained 8A8-containing fat (triglyceride) by a conventional high performance liquid chromatography.

Example 3 FIG. Evaluation of learning ability of TGA40S by Morris-type water maze test As an experimental group of old rats, 16 18-month-old male Fischer rats were used as a control feed group (8 animals: OC group) and a TGA40S combination feed group (8 animals: OA group). The control feed and the SUNTGA40S combination feed shown in Table 1 were given to each group. Then, as a control group (YC group) of young rats, the control diet shown in Table 1 was given to 8 4-month-old male Fischer rats (YC group). In addition, what was obtained in Example 1 was used for TGA40S used for the TGA40S mixing feed.

Table 1 Experimental food ─────────────────────────────
Control feed with TGA40S
(G) (g)
─────────────────────────────
Casein 200 200
DL-methionine 3 3
Corn starch 150 150
Sucrose 500 500
Cellulose powder 50 50
Corn oil 50 45
Minerals AIN-76 35 35
Vitamins AIN-76 10 10
Choline bitartrate 2 2
Vitamin E 0.05 0.05
TGA40S 0 5
─────────────────────────────

  Since the daily food intake per rat is about 20 g, the daily intake per rat of TGA40S is 0.1 g. Since 40% by mass of all fatty acids bound to TGA40S is arachidonic acid, the daily intake of arachidonic acid per rat is 40 mg (ignoring the mass of the glycerol skeleton for convenience) Calculation). This 40 mg is equivalent to 133 mg / 60 kg / day in terms of human intake.

  A Morris water maze learning test was conducted before and after the third month of breeding (21 months for old rats and 7 months for young rats). Morris-type water maze test is a water tank (120cm in diameter, 35cm in height) with black ink turbid water (20cm in height), and an escape platform (11.5cm in diameter, large enough for a rat to stand up). 19cm high) (the escape platform is below the surface of the water, and the rats that swim in the tank cannot see the escape platform), and put the rat to be trained from a predetermined position in this tank (starting point) In a learning test based on spatial recognition that allows you to swim to the escape platform, it is recognized for its connection with the brain hippocampus that controls memory, and is widely used in Europe and the United States. A schematic illustration of an apparatus used for the Morris water maze test is shown in FIG.

  As the number of times is increased, the rat learns the position of the escape platform. Rats were trained as follows. That is, if the rat is released from the starting point of the Morris water maze test apparatus and the rat cannot reach the escape platform even after 60 seconds, the rat is placed on the escape platform and the invisible escape platform The position of was learned. This study was continued for two weeks, limited to twice a day. The ratio of time spent swimming within a range of ± 15 ° C. from the starting point to the escape platform from the starting point to the total swimming time (Hit%, see FIG. 2) was used as a learning index. Compared to young rats, the acquisition rate of learning in old rats was clearly reduced, but by giving TAG40S, that is, arachidonic acid, it approached and improved the level of young rats (FIG. 3). In FIG. 3, the horizontal axis represents 4 trials, that is, 2 days as one scale.

  Next, in order to measure the degree of learning acquisition, a probe test was performed on the next day, ie, the 15th day, after the two weeks of learning. After learning, when the escape platform is removed, the rat swims around where it was previously. Memorize the location of the escape platform and swim around the place (evaluated by the time (seconds) spent in the quarter (1/4) with the aquarium divided into 4 sections) Thus, the degree of learning acquisition can be evaluated. The swimming trajectories of No. 1 and No. 2 rats in YC group, OA group and OC group are shown (FIG. 4). Since the rats were trained by changing the starting point depending on the individual, the starting point (S) and the section with the escape platform in FIG. 4 differ depending on the individual of the rat. In FIG. 5, target quadrant represents a section (1/4) where the escape platform is located. Rat OC-1 in the OC group was clearly strayed, and only 2.4 seconds were in the compartment with the escape platform. The results of the probe test are summarized in Table 2.

であり、ここで、

は平均(各群の平均)、μは母平均（１５秒)、Ｓは標本標準偏差、ＳＤは平均±標準偏差、ｎはデータ数（各群のデータ数）を表し、Ｓは、

を表し、ＳＤは、

を表す。 In Table 2,

And where

Is the mean (average of each group), μ is the population mean (15 seconds), S is the sample standard deviation, SD is the mean ± standard deviation, n is the number of data (number of data in each group), S is

And SD is

Represents.

  When Table 2 is shown in a graph (FIG. 5), for the OA group given TGA40S to old rats, the time spent in the section where the escape platform was located (the location of the escape platform was remembered and the place where it was swimming) (Time) is significantly longer. The chance level of 15 seconds indicates the possibility of staying by chance because the residence time is measured by swimming the rat for 60 seconds. The bar graph shows the average time spent in the compartment where the rat's escape platform was located.

  Next, the brain hippocampus was excised from the rat subjected to the Morris water maze test, and the total lipid was extracted by the Folch method. Then, fractionate lipids by thin layer chromatography, scrape phospholipid fractions, remove water by azeotropy with ethanol, and analyze by fatty acid methyl ester with 10% hydrochloric acid-methanol by gas chromatography. did. Correlation of water maze learning parameters ("arrival time to escape platform (shorter is better)" "ratio of swimming toward escape platform (Hit%, larger is better)") and the amount of arachidonic acid in the hippocampus As a result of the first-order approximation curve based on the least squares method (Fig. 6), in the time to reach the escape platform, the amount of hippocampal arachidonic acid was negatively correlated (correlation coefficient R = -0.38). A positive correlation (correlation coefficient R = 0.32) was observed. In FIG. 6, the vertical axis represents mg of arachidonic acid per 1 g of hippocampal tissue. Thus, it was revealed for the first time that TGA40S improves learning ability or cognitive ability, and it has been proved for the first time that the effect is due to arachidonic acid.

Example 4 Evaluation of learning ability of 8A8 by Morris-type water maze test As an experimental group of old rats, 20 18-month-old male Fischer rats were used as a control feed group (6 animals: OC group), and TGA40S-containing food group (6 animals: OA group). And 8A8 combination feed group (8 animals: 8A8 group), and the control feed, TGA40S combination feed and 8A8 combination feed shown in Table 3 were given to each group. Then, as a control group (YC group) of young rats, the control diet shown in Table 3 was given to 8 4-month-old male Fischer rats. In addition, 96 mol% 8A8 obtained in Example 2 was used for 8A8 used for 8A8 combination feed.

Table 3 Experimental food ────────────────────────────────────
Control feed TGA40SS formula feed 8A8 formula feed
(G) (g) (g)
────────────────────────────────────
Casein 200 200 200
DL-methionine 3 3 3
Cornstarch 150 150 150
Sucrose 500 500 500
Cellulose powder 50 50 50
Corn oil 50 45 45.8
Minerals AIN-76 35 35 35
Vitamins AIN-76 10 10 10
Choline bitartrate 2 2 2
Vitamin E 0.05 0.05 0.05
TGA40S 0 5 0
8A8 0 0 4.2
────────────────────────────────────

  Since the daily food intake per rat is about 20 g, the daily intake per rat of TGA40S is 0.1 g. Since 40% by mass of all fatty acids bound to TGA40S is arachidonic acid, the daily intake of arachidonic acid per rat is 40 mg (calculated by ignoring the mass of the glycerol skeleton for convenience) . Since the molecular weight of TGA40S is 928.5 (calculated from the average fatty acid molecular weight) and the molecular weight of 8A8 is 628.7, the 8A8 mixed feed group is also tested so that the daily intake of arachidonic acid per rat is 40 mg. The food was designed.

A Morris water maze learning test was conducted before and after the third month of breeding (21 months for old rats and 7 months for young rats).
As a result of the probe test, the position of the escape platform was memorized, and the time spent swimming around the place (the time when the aquarium was divided into 4 sections and the section (1/4) where the escape platform was located (seconds) ) Was 28.59 ± 5.44 ^a , 13.27 ± 7.89 ^b , 22.02 ± 5.35 ^c and 27.18 ± 5.10 ^ac in the YC group, OC group, OA group and 8A8 group (numerical value: standard value ± standard deviation) , A, b, and c are significantly different between different characters (P <0.05)). Therefore, by providing triglycerides containing arachidonic acid as a constituent fatty acid, the degree of learning that decreased with aging was significantly improved toward the level of young rats. In comparison between TGA40S and 8A8, 8A8 showed a tendency of higher learning acquisition. Both OA group and 8A8 group have the same arachidonic acid intake in rats, indicating that 8A8 is more easily absorbed and proved to be effective for pancreatic lipase whose activity decreased with age. did.

Embodiment 5 FIG. Evaluation of learning ability of triglycerides containing 5% or more of 8A8 by Morris-type water maze test As an experimental group of old rats, 20 18-month-old male Fischer rats were used as a control diet group (6 animals: OC group), 8A8-containing diet group ( 6 animals: 8A8 group) and 8A8-containing fat and oil-containing feed group (8 animals: 8A8 (32 mol%) group), and each group contains the control feed, 8A8-containing feed and 8A8 An oil and fat mixed feed was given. Then, as a control group of young rats, the control diets shown in Table 4 were given to 8 4-month-old male Fischer rats (YC group). The 8A8 used in the 8A8-containing feed contains 96 mol% 8A8 obtained in Example 2, and the 8A8-containing fat (triglyceride) used in the 8A8-containing fat-containing feed contains 31.6 mol% of 8A8 obtained in Example 2. Oil (triglyceride) was used.

Table 4 Experimental food ────────────────────────────────────
Control feed 8A8-containing feed 8A8-containing fat-containing feed
(G) (g) (g)
────────────────────────────────────
Casein 200 200 200
DL-methionine 3 3 3
Cornstarch 150 150 150
Sucrose 500 500 500
Cellulose powder 50 50 50
Corn oil 50 45.8 45.8
Minerals AIN-76 35 35 35
Vitamins AIN-76 10 10 10
Choline bitartrate 2 2 2
Vitamin E 0.05 0.05 0.05
8A8 0 4.2 0
Oil containing 8A8 0 0 4.2
────────────────────────────────────

The 8A8 mixed feed is the same as in Example 4, and the daily intake of arachidonic acid per rat is 40 mg. In the case of a diet containing 8A8-containing fats and oils (triglycerides), the daily intake of arachidonic acid per rat is 13.2 mg.
A Morris water maze learning test was conducted before and after the third month of breeding (21 months for old rats and 7 months for young rats).

As a result of the probe test, the position of the escape platform was memorized, and the time spent swimming around the place (the time when the aquarium was divided into 4 sections and the section (1/4) where the escape platform was located (seconds) Was evaluated as 27.91 ± 5.93 ^a , 13.75 ± 7.74 ^b , 27.00 ± 4.65 ^a and 21.18 ± 4.89 ^c in the YC group, OC group, 8A8 group and 8A8 (32% mol) group, respectively (numerical values: Mean ± standard deviation, a, b, and c are significantly different between different characters (P <0.05)). Therefore, by providing an oil (triglyceride) containing 5 mol% or more of 8A8, the degree of learning that decreased with aging was significantly improved toward the level of young rats. However, the degree of acquisition was clearly lower than that of the 8A8 mixed feed group, demonstrating that it depends on the concentration of 8A8 and further on the concentration of arachidonic acid.

Example 6 Preparation of capsule containing oil and fat (triglyceride) containing arachidonic acid Water was added to 100 parts by mass of gelatin and 35 parts by mass of glycerin for food additives, and dissolved at 50-60 ° C. to prepare a gelatin coating having a viscosity of 2000 cp. Next, 0.05 mass% of vitamin E oil was mixed with the arachidonic acid-containing fat (triglyceride) obtained in Example 1 to prepare a content 1. Content 2 was prepared by blending 0.05% by mass of vitamin E oil with the oil (triglyceride) containing 32 mol% of 8A8 obtained in Example 2.

  50% by mass of oil (triglyceride) containing arachidonic acid obtained in Example 1 and 50% by mass of fish oil (tuna oil: the proportions of eicosapentaenoic acid and docosahexaenoic acid in the total fatty acids are 5.1% by mass and 26.5% by mass, respectively) Then, content 3 was prepared by mixing 0.05% by mass of vitamin E oil. 80% arachidonic acid-containing fat (triglyceride) obtained in Example 1 and 20% by weight of fish oil (tuna oil: the proportions of eicosapentaenoic acid and docosahexaenoic acid in the total fatty acids are 5.1% and 26.5% by mass, respectively) The content 4 was prepared by mixing 0.05% by mass of vitamin E oil. Using these contents 1 to 4, capsules were formed and dried by a conventional method to produce soft capsules containing 180 mg of contents per grain.

Example 7 Use in fat infusions After adding 400 g of oil (triglyceride) containing 32 mol% of 8A8 obtained in Example 2, 48 g of purified egg yolk lecithin, 20 g of oleic acid, 100 g of glycerin and 40 ml of 0.1N caustic soda, and then dispersing with a homogenizer Add distilled water for injection to make 4 liters. This was emulsified with a high-pressure spray type emulsifier to prepare a lipid emulsion. After 200 ml of the lipid emulsion is dispensed into a plastic bag, it is autoclaved at 121 ° C. for 20 minutes to obtain a fat infusion.

Example 8 FIG. Use in juice Add 2 g of β-cyclodextrin to 20 ml of 20% ethanol aqueous solution and add 100 mg of arachidonic acid-containing triglyceride (containing 0.05 mass% vitamin E) obtained in Example 1 while stirring with a stirrer. Incubated at 50 ° C. for 2 hours. After cooling at room temperature (about 1 hour), the mixture was further incubated at 4 ° C. for 10 hours while continuing to stir. The produced precipitate was collected by centrifugation, washed with n-hexane, and lyophilized to obtain 1.8 g of a cyclodextrin inclusion compound containing arachidonic acid-containing triglyceride. 1 g of this powder was uniformly mixed with 10 L of juice to prepare a juice containing arachidonic acid-containing triglycerides.

Effects of the Invention According to the present invention, a symptom or disease preventive or ameliorating agent caused by a decrease in brain function, a symptom caused by a decrease in brain function, or an arachidonic acid or a compound comprising arachidonic acid as a constituent fatty acid as an active ingredient It is possible to provide a food and drink having a disease prevention or ameliorating action and a method for producing the same, and is particularly useful for human beings toward an aging society.

Claims (9)

( B) containing only arachidonic acid and / or (b) an alcohol ester of arachidonic acid or a compound in which some or all of the constituent fatty acids are triglycerides which are arachidonic acid or arachidonic acid which is a glycolipid as a constituent fatty acid. A pharmaceutical composition for preventing or improving a decrease in memory / learning ability. The pharmaceutical composition according to claim 1 , wherein the triglyceride in which some or all of the constituent fatty acids are arachidonic acid is a triglyceride in which a medium chain fatty acid is bonded to the 1,3-position and arachidonic acid is bonded to the 2-position. The pharmaceutical composition according to claim 2 , wherein the medium chain fatty acid is selected from fatty acids having 6 to 12 carbon atoms. A pharmaceutical composition for preventing or improving a decrease in memory / learning ability, comprising as an active ingredient only a triglyceride in which a part or all of the constituent fatty acid is arachidonic acid. Some or all of the constituent fatty acids of the triglycerides containing triglycerides is arachidonic acid, the proportion of arachidonic acid, characterized in that 10 mass% or more with respect to total fatty acids constituting the triglyceride, to claim 4 The pharmaceutical composition as described. The pharmaceutical composition according to claim 4 or 5 , wherein the triglyceride containing a triglyceride in which a part or all of the constituent fatty acid is arachidonic acid is extracted from a microorganism belonging to the genus Mortierella. Some or all of the constituent fatty acids triglycerides containing triglycerides is arachidonic acid, a pharmaceutical composition according to any one of claims 1 to 6 eicosapentaenoic acid is that does not contain triglycerides. Prevention of symptoms or diseases caused by reduced brain function, containing only triglycerides containing 5 mol% or more of triglycerides with medium chain fatty acids at the 1,3-position and arachidonic acid at the 2-position as active ingredients Or a pharmaceutical composition having an improving action. The pharmaceutical composition according to claim 8 , wherein the medium chain fatty acid is selected from fatty acids having 6 to 12 carbon atoms.

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