Oral composition for enhancing expression or production of ciliary neurotrophic factor or improving cognitive function

Abstract

The present invention provides an oral composition having the action of enhancing the expression of a ciliary neurotrophic factor (CNTF), in particular, the expression thereof in the hippocampus, or an oral composition having the action of improving cognitive function. Specifically, provided is an oral composition for enhancing expression or promoting the production of a CNTF, or for improving cognitive function, the composition containing, as an active ingredient, white mold cheese or a processed product thereof.

Description

[Technical Field]

[0001] The present disclosure relates to an oral composition that enhances the expression of ciliary neurotrophic factor (CNTF) (hereinafter also referred to simply as "CNTF" in the present invention), particularly in the hippocampus of a mammal. The present disclosure also relates to an oral composition that improves cognitive function in a mammal. [Background technology]

[0002] The Ministry of Health, Labor and Welfare announced in 2012 that the estimated number of dementia patients and mild cognitive impairment (MCI) patients in Japan is 4.62 million and 4 million, respectively, and these rates are expected to increase year by year (Non-Patent Document 1). Dementia is a major cause of impairment in activities of daily living (ADL), which are basic activities essential for living (eating, dressing, moving, toileting, grooming, bathing, etc.) (Non-Patent Document 2), and significantly reduces the quality of life of elderly people and their caregivers, such as their families. Therefore, preventing the onset of dementia is a socially important issue.

[0003] Large-scale epidemiological studies have shown that the consumption of milk and dairy products is a protective factor against dementia (Non-Patent Document 3), but details are unclear. It has also been reported that certain peptides derived from milk proteins can enhance memory, learning, and / or cognitive function (Patent Documents 1 to 4). Furthermore, although the results were reported for an Alzheimer's model animal (mouse) created by genetic mutation, it is known that the consumption of Camembert cheese improves cognitive function and increases the expression of brain-derived neurotropic factor (BDNF) (hereinafter simply referred to as "BDNF") and glial cell line-derived neurotrophic factor (GDNF) (hereinafter simply referred to as "GDNF") (Non-Patent Document 4). Furthermore, exercise increases BDNF and improves performance in areas such as memory and learning, suggesting a relationship between BDNF and cognitive function (Non-Patent Document 5).

[0004] Neurotrophic factors, in the narrow sense, are survival factors required by neurons. They are produced by tissues to which neurons project and function to maintain the survival of neurons. There are several neurotrophic factors, including the aforementioned BDNF and GDNF. They are broadly classified into three types: target-derived neurotrophic factors, pathway-derived factors, and autocrine factors. Target-derived neurotrophic factors are produced by neurons or glia near the nerve terminals of nerve cells, taken up by receptors in the nerve terminals, and then transported to the cell bodies of target neurons via retrograde transport. Nerve growth factor (NGF) is a representative example of such factors. Pathway-derived factors are a group of factors produced by oligodendrocytes and astroglia surrounding the axons of neurons, and are taken up by the axons into neurons to act. A representative example of these is ciliary neurotrophic factor (CNTF). Autocrine factors are a group of factors produced in the neuronal cell body, taken up by receptors in the neuronal cell body itself or in nearby neuronal cell bodies, and act on the neuronal cell body. Representative examples of these include acidic fibroblast growth factor (aFGF) and insulin-like growth factor (IGF) (all from Non-Patent Document 6).

[0005] Among these neurotrophic factors, CNTF, a pathway-derived factor, was initially identified as a survival factor acting on neurons emanating from the ciliary ganglion of chick embryos. However, it has recently been shown to promote the survival and differentiation of nervous system neurons, including motor neurons, sympathetic ganglion neurons, sensory neurons, hippocampal neurons, and medial septal neurons. It is also known to prevent degeneration of motor neurons after axotomy and promote astrocyte differentiation and oligodendrocyte survival and maturation (Non-Patent Document 7). Furthermore, Non-Patent Document 8 suggests that CNTF, due to its ability to prevent neuronal apoptosis, may improve cognitive function and prevent Alzheimer's disease. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Publication No. 2017-008104 [Patent Document 2] Japanese Patent Application Laid-Open No. 2012-012358 [Patent Document 3] Japanese Patent Application Laid-Open No. 2015-154773 [Patent Document 4] Japanese Patent Application Laid-Open No. 2012-031139 [Non-patent literature]

[0007] [Non-Patent Document 1] Asada T. et al., 2013, Prevalence of dementia in urban areas and responses to dementia-related functional impairments. Ministry of Health, Labour and Welfare Science Research Grant, Comprehensive Research Project on Dementia Prevention, FY2011-FY2012 Comprehensive Research Report [Non-patent document 2] Yoshida D et al. 2012, J Epidemiol. 22: 222-229. [Non-patent document 3] Ozawa M et al., 2014, J Am Geriatr Soc., 62:1224-1230. [Non-patent document 4] Ano Y et al., 2015, PLOS One, 10:e0118512 [Non-patent document 5] Neeper SA et al., 1996, Brain Res., 726:49-56 [Non-patent document 6] Kazuo Yamada, "Gene Expression and Receptor Mechanism of Neurotrophic Factors," Neurosurgery, Vol. 5, No. 2, pp. 118-127, March 1996 [Non-Patent Document 7] Sigma-Aldrich webpage: "Ciliary Neurotrophic Factor Human" (https: / / www.sigmaaldrich.com / catalog / product / sigma / c3710?lang=ja®ion=JP) [Non-patent document 8] Pierre Garcia et al., The Journal of Neuroscience, June 2, 2010, 30(22):7516-7527 [Non-Patent Document 9] "Modern Cheese Studies" by Keiji Ikoshi, Food Materials Research Association, pp. 28-30, February 2011 Summary of the Invention [Problem to be solved by the invention]

[0008] The present disclosure aims to provide an oral composition that has the effect of enhancing ciliary neurotrophic factor (CNTF) expression, particularly in the hippocampus of a mammal, and also has the effect of improving cognitive function in a mammal. [Means for solving the problem]

[0009] To address the above-mentioned issues, the present inventors conducted extensive research using mice with cognitive decline due to a high-fat diet. They found that oral administration of white mold cheese to these mice dose-dependently increased CNTF gene (mRNA) expression in the hippocampus. Furthermore, they confirmed that the decline in episodic memory induced by a high-fat diet was suppressed and improved. Based on these findings, the present inventors believe that the ingestion of white mold cheese enhances CNTF expression and promotes the survival and differentiation of neurons, particularly hippocampal neurons. Furthermore, the present inventors confirmed that the content of oleic acid amide and palmitic acid amide in white mold cheese significantly increased after heat treatment. These fatty acid amides are known to inhibit the accumulation of amyloid and tau proteins, which are associated with brain aging. Therefore, it is believed that heat treatment of white mold cheese can further enhance these effects. The present disclosure was completed based on this finding and includes the following embodiments.

[0010] (I) Oral composition for enhancing expression or production of CNTF or improving cognitive function (I-1) An oral composition for enhancing the expression or production of CNTF or improving cognitive function, comprising white mold cheese or a processed product thereof as an active ingredient. (I-2) The oral composition described in (I-1), wherein the white mold cheese is Camembert cheese. (I-3) An oral composition according to (I-1) or (I-2), wherein the white mold cheese is heat-treated. (I-4) An oral composition according to any one of (I-1) to (I-3), which is a food composition or a food additive composition. (I-5) The oral composition according to (I-4), which is a health food, functional food, nutritional supplement, supplement, health functional food, food for specified health uses, nutritional functional food, food with functional claims, or food for patients.

[0011] (II) A method for producing an oral composition for enhancing the expression or production of CNTF or improving cognitive function (II-1) A method for producing an oral composition for enhancing the expression or production of CNTF or improving cognitive function, comprising a step of heat-treating white mold cheese. (II-2) A manufacturing method described in (II-1), in which the heat treatment step is a step of increasing the content of at least one fatty acid amide selected from the group consisting of oleic acid amide and palmitic acid amide in white mold cheese. (II-3) The manufacturing method described in (II-1) or (II-2), wherein the white mold cheese is Camembert cheese. (II-4) A method for producing an oral composition for enhancing the expression or production of CNTF or improving cognitive function, characterized in that white mold cheese or a processed product thereof is blended into the raw material composition of the oral composition.

[0012] (III) Method for increasing fatty acid amides (III-1) A method for increasing the content of at least one fatty acid amide selected from the group consisting of oleic acid amide and palmitic acid amide in white mold cheese, comprising a step of heat-treating the white mold cheese. (III-2) The method described in (III-1), wherein the white mold cheese is Camembert cheese.

[0013] (IV) Uses of white mold cheese or its processed products (IV-1) Use of white mold cheese or a processed product thereof for producing an oral composition for enhancing the expression or promoting the production of CNTF in the hippocampus of a mammal, or an oral composition for improving the cognitive function of a mammal. (IV-2) The use described in (IV-1), wherein the white mold cheese is heat-treated. (IV-3) The use according to (IV-1) or (IV-2), wherein the white mold cheese is Camembert cheese. (IV-4) A method for enhancing the expression or production of CNTF in the hippocampus or improving cognitive function in a subject suffering from cognitive decline or a disease or symptom resulting therefrom, the method comprising orally administering heat-treated white mold cheese or a processed product thereof. (IV-5) The method described in (IV-4), wherein the cognitive function is a memory-related cognitive function. (IV-6) The method according to (IV-4) or (IV-5), wherein the cognitive decline is due to high-fat diet intake or aging. [Effects of the Invention]

[0014] According to the present disclosure, an oral composition is provided that has the effect of enhancing CNTF expression, particularly in the mammalian hippocampus. By ingesting the oral composition, various physiological effects (e.g., the effect of maintaining the survival or promoting the differentiation of nervous system neurons) and the physiological functions derived therefrom can be enjoyed, which are associated with enhanced CNTF expression or enhanced CNTF production in the hippocampus.

[0015] Furthermore, according to the present disclosure, the content of at least one fatty acid amide selected from the group consisting of oleic acid amide and palmitic acid amide in white mold cheese can be increased. White mold cheese with an increased fatty acid amide content is effective in improving cognitive decline in mammals. In particular, it can be effectively used to improve cognitive decline related to memory. [Brief explanation of the drawings]

[0016] [Figure 1] The results of the hippocampal ciliary neurotrophic factor (CNTF) mRNA expression assay performed in Experimental Example 1(1) are shown. In the figure, "Cont" refers to Group 1 (control group), "LC" refers to Group 2 (low-dose Camembert (8 g / kg / day) administration group), and "HC" refers to Group 3 (high-dose Camembert (15 g / kg / day) administration group) (same as in Figure 2). [Figure 2](A) A diagram showing the details (habituation trial [Habituation], acquisition trial [Trial 1], test trial [Trial 2]) of the behavioral test (Object Recognition Test) conducted in Experimental Example 1(2). (B) The results of the behavioral test (ORT) conducted in Experimental Example 1(2) are shown. The results show the ratio (%) of approach time to the novel object to the total approach time to two objects placed in a rectangular open field (vertical axis: Novel object / Total exploration (%)). [Figure 3] 1 shows the experimental results of Experimental Example 3, which show the relationship between the heating conditions (temperature, time) of white mold cheese and the oleic acid amide content (μg / g) in the white mold cheese. DETAILED DESCRIPTION OF THE INVENTION

[0017] (I) Oral composition for enhancing expression or production of CNTF or improving cognitive function The oral composition of the present invention for enhancing the expression or promoting the production of CNTF or for improving cognitive function (hereinafter, when referring to these collectively, simply referred to as "the oral composition") is characterized by containing white mold cheese as an active ingredient.

[0018] In the present disclosure, "enhancing CNTF expression" refers to enhancing CNTF mRNA expression in a mammal. This includes suppressing or preventing a decrease in CNTF expression, maintaining the amount of CNTF in the body by suppressing a decrease in CNTF expression, and enhancing or improving CNTF expression so as to prevent (or maintain) a decrease in CNTF amount. Furthermore, enhanced CNTF mRNA expression results in promotion of CNTF production. Therefore, in the present invention, "promoting CNTF production" includes suppressing a decrease in CNTF mRNA expression in a mammal, and maintaining the amount of CNTF in the body. The hippocampus is a preferred example of a mammalian body.

[0019] Enhanced CNTF expression can maintain or promote the survival or differentiation of various neurons involved in CNTF (e.g., motor neurons, sympathetic ganglion neurons, sensory neurons, hippocampal neurons, medial septal neurons, etc.), and is thought to be particularly effective in maintaining or promoting the survival or differentiation of hippocampal neurons.

[0020] Furthermore, as shown in the experimental examples described below, oral administration of this oral composition has been shown to improve impaired cognitive function. This suggests that the aforementioned increase in CNTF expression in the hippocampus may contribute to the improvement of cognitive function. Therefore, this oral composition is expected to improve cognitive function as a result of increasing CNTF expression.

[0021] Here, "cognitive function" refers to cognitive function, particularly memory-related cognitive function, as assessed by the Object Recognition Test (ORT), a behavioral test known to assess cognitive function in mammals. "Improving cognitive function" refers to improving cognitive function, particularly memory-related brain function, and includes suppressing or preventing cognitive function decline, maintaining cognitive function by suppressing cognitive function decline, and improving cognitive function to prevent (or maintain) cognitive function decline. Improvement of cognitive function also includes, for example, recovery of cognitive ability that has declined or symptoms that indicate decline. The "cognitive function" also includes memory and learning function, to the extent described above. The cause of cognitive decline is not important, but includes, for example, cognitive decline due to high-fat diet intake and cognitive decline associated with aging. As mentioned above, oleic acid amide and / or palmitic acid amide are known to have the effect of suppressing the accumulation of amyloid and tau proteins associated with brain aging. As shown in Experimental Examples 2 and 3 described below, the oleic acid amide content and / or palmitic acid amide content can be significantly increased by heat-treating white mold cheese. Therefore, it is considered that the use of heat-treated white mold cheese as a raw material for the oral composition can further enhance its cognitive function improving effect.

[0022] The white mold cheese used as an active ingredient in this oral composition is a type of natural cheese as defined in the "Ministerial Ordinance on the Ingredient Standards of Milk and Dairy Products" (Ministry of Health and Welfare Ordinance No. 52 of December 27, 1951). According to "Modern Cheese Studies" (Keiji Ikoshi, Food Materials Research Association, pp. 28-30, February 2011) (Non-Patent Document 9), natural cheeses are classified into seven types: (1) fresh type (unaged cheese), (2) white mold type (soft cheese with a white mold rind), (3) washed type (soft cheese in which special microorganisms are planted on the cheese rind (some naturally occur) and washed with salt water or local sake), (4) chèvre type (cheese made from goat's milk), (5) blue mold type (so-called blue cheese, in which blue mold is allowed to grow and its action creates flavor from within), (6) semi-hard type (cheese characterized by a low moisture content and a firm, elastic texture), and (7) hard type (cheese that is the hardest of all cheeses, aged for six months to at most two years or more, and can withstand long-term storage). White mold cheese falls into the category of (2) white mold type.

[0023] White mold cheese has a soft cheese body and a white mold layer covering at least a portion of the surface of the soft cheese body. The soft cheese body is in a matured (fermented) state due to the white mold contained in the white mold layer. The white mold layer may cover a portion of the surface of the soft cheese body, or may cover the entire surface of the soft cheese body. In an embodiment in which the white mold layer covers a portion of the surface of the soft cheese body, the proportion of the area of ​​the portion covered by the white mold layer to the surface area of ​​the soft cheese body is not limited, and can be, for example, in the range of 30 to 100%. Examples of white mold contained in the white mold layer include Penicillium camemberti, Penicillium candidum, and Penicillium caseicolum. The white mold contained in the white mold layer may be one type, or two or more types.

[0024] White mold cheeses include Camembert, Brie, Padaffey, Brique de Bache, Baccarat, double cream type Supreme, Caprice des Dieu, triple cream type Saint-André, white Castello, and Cambozola, which has a white mold surface and a blue mold center. Also included are types in which spices such as pepper and herbs are mixed into white mold cheese, or types in which the cheese is soaked in spices in brine or coated with spices. While not limited to these, Camembert cheese is preferred.

[0025] White mold cheese can be produced according to a conventional method for producing white mold cheese, for example, Camembert cheese, by the following method. Raw milk is prepared using raw milk, skim milk, partially skim milk, cream, or the like, and pasteurized as necessary. The raw milk is heated, and lactic acid bacteria and rennet (a milk-clotting enzyme) are added. Whey is removed from the obtained curd to obtain cheese curds. The cheese curds are placed in a mold and shaped. The shaped cheese curds are salted, and white mold is sprayed onto the surface of the cheese curds. The cheese curds with the white mold sprayed onto the surface are aged in a temperature- and humidity-controlled aging chamber. Examples of white molds used in producing white mold cheese include the above-mentioned white molds belonging to the Penicillium genus. The white molds may be of one type or two or more types. The aging period is usually 1 to 4 weeks, with a longer aging period of 2 to 4 weeks being preferred to maximize the effects of the present invention.

[0026] In the production of white mold cheese, primary and secondary degradation occur during the ripening period. Examples of primary degradation include the production of amino acids through the degradation of casein and the production of fatty acids through the degradation of milk fat. Examples of secondary degradation include the production of aroma substances through the degradation of primary degradation products such as amino acids and fatty acids. Primary degradation is generally common to all ripened natural cheeses. The flavor unique to each natural cheese is mainly imparted by secondary degradation. In the production of white mold cheese, white mold grows on the surface of the cheese curd during the ripening period, and the amino acid dehydrogenase contained in the white mold decomposes amino acids to produce ammonia. The ammonia produced on the surface of the cheese curd increases the pH outside the cheese curd. The resulting pH gradient migrates calcium from the inside of the cheese curd to the outside, promoting the softening of the tissue inside the cheese curd. The pH of the entire white mold cheese at the completion of ripening is, for example, pH 4.6 to 7. A pH of 5 to 7 is preferable to maximize the effects of the present invention.

[0027] The white mold cheese used in the present disclosure may be heat-treated after being produced by the above-mentioned conventional method (after completion of aging). In other words, the white mold cheese may be either unheat-treated or heat-treated. As shown in the experimental examples described below, heat treatment increases the content of oleic acid amide and / or palmitic acid amide (hereinafter, these are also collectively referred to as "fatty acid amides") in the white mold cheese, and the effects of the present disclosure are expected to be enhanced and more pronounced, so heat-treated cheese is preferred.

[0028] The heat treatment step may be carried out after aging, for example, after the white mold cheese is cut into portions, packaged, or filled into containers. The heat treatment of white mold cheese may also serve as a sterilization treatment, in which case general heat sterilization conditions, sterilization methods, sterilization equipment, etc. used for sterilization, such as retort sterilization, can be used as is.

[0029] The heat treatment temperature should be such that the central temperature of the cheese is 60°C or higher, since the fatty acid amide content increases with increasing temperature. A temperature that results in a central temperature of 130°C or lower is preferred. A central temperature of 70°C or higher and 120°C or lower is more preferred, and from the viewpoint of sterilizing microorganisms such as mold, a temperature of 80°C or higher and 110°C or lower is preferred. The heat treatment time can be adjusted according to the heat treatment temperature. A shorter time is sufficient if the heat treatment temperature is high, but the same effect can be achieved by heating for a longer time if the heat treatment temperature is low. For example, the heat treatment time can be 10 minutes or higher and 120 minutes or lower. A more preferred time is 20 minutes or higher and 90 minutes or lower. Specifically, the cheese may be kept at a core temperature of 80°C or higher for 10 minutes or longer. More preferably, the cheese may be kept at a core temperature of 80°C or higher for 20 minutes or longer. Preferred conditions for increasing the fatty acid amide content include keeping the cheese at 95°C or higher for 30 minutes or longer, and more preferably keeping the cheese at 120°C for 30 minutes or longer.

[0030] Such heat treatment can increase the fatty acid amide content in white mold cheese, thereby improving cognitive function, and also imparts a good flavor to the white mold cheese and maintains its quality for a long period of time. Furthermore, heat treatment after aging is also preferable in that it reduces the risk of secondary bacterial contamination and improves shelf life and safety. If the white mold cheese is to be heat-treated after being cut into portions and packaged, it is preferable to provide a thin film of thickening polysaccharides, gelatin, oils or fats, cream, etc. on the inside of the packaging. This will prevent the cheese from sticking to the packaging after heating, making it easier to remove. The white mold cheese can be frozen after such heat treatment or after being packaged after heat treatment, and can be stored or distributed in the market in a frozen state.

[0031] As the active ingredient of the oral composition, a processed white mold cheese can be used in place of or in combination with the white mold cheese produced by the above-mentioned method. Here, processed products include those that are different in form from the above-mentioned white mold cheese but are considered to be the same in terms of ingredients. For example, processed white mold cheese includes pulverized products obtained by drying and pulverizing white mold cheese, and pastes obtained by semi-fluidizing white mold cheese. The white mold cheese may be heat-treated or unheat-treated. These pulverized products or pastes can be considered identical to white mold cheese in terms of ingredients, since they contain milk components derived from the soft cheese body of white mold cheese and components from the white mold layer.

[0032] The dosage of the active ingredient, white mold cheese, of the oral composition is not particularly limited as long as it demonstrates the effects of the present invention. However, taking into account the results obtained in the Examples described below, the lower limit of the dosage may be, for example, 10 g or more, 15 g or more, 20 g or more, 30 g or more, 40 g or more, or 50 g or more per day. The upper limit of the dosage may be, for example, 100 g or less, 90 g or less, 80 g or less, 70 g or less, or 60 g or less per day. Furthermore, the range of the dosage may be, for example, 10 g or more to 60 g or 15 g or more to 50 g.

[0033] (II) Application to food or food additive compositions The use forms of the oral composition include food compositions and food additive compositions. Here, the term "food composition" refers to a composition that is consumed directly without being mixed with other ingredients. The term "food additive composition" refers to a composition that is added to other foods and beverages when consumed, or to other food and beverage ingredients during food production. Both have in common the fact that they are used as foods and beverages. Such oral compositions may consist solely of the aforementioned white mold cheese and / or processed products thereof. Furthermore, they may contain other ingredients in addition to the white mold cheese and / or processed products thereof, provided that the effects of the present invention are not impaired. Examples of other ingredients include ingredients consumed as food and raw ingredients used in food production, including additives used in food production. Here, the term "food" is synonymous with "food and drink" and is used in a broad sense to encompass beverages. Furthermore, the term "food" as used in this disclosure includes not only food for humans but also food for animals such as pets and livestock (i.e., including feed and laboratory feed).

[0034] The form of the food composition and food additive composition is not particularly limited, and may be in the form of the white mold cheese described above, or in any other form such as solid, powder, granules, semi-liquid (including paste), and also includes forms such as beverages.

[0035] The food composition also includes so-called health foods, functional foods, nutritional supplements, and supplements, including health functional foods (foods for specified health uses, nutritional functional foods, and functional foods) and foods for patients, such as foods labeled for disease risk reduction. Supplements can be in the form of tablets produced by adding excipients, binders, etc. to processed products such as the dried and pulverized Camembert cheese, kneading the mixture, and then tableting. They can also be in the form of powders, pills, capsules, jellies, granules, etc. The amount of the processed white mold cheese to be added is not particularly limited, as long as it is an effective amount that produces the effects of the present invention.

[0036] These supplements can also be formulated in combination with the following ingredients and compounds that are said to have the effect of improving brain function: These include food ingredients such as ginkgo biloba extract, arachidonic acid (ARA), GABA, theanine, ceramide, caffeine, carnitine, α-glycerylphosphorylcholine (α-GPC), Bacopa monniera, DHA-binding phospholipids, phosphatidylserine (PS), phosphatidylcholine, St. John's wort, astaxanthin, niacin, pyrroloquinoline quinone (PQQ), and coenzyme Q10 (CoQ10); unsaturated fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA); polyphenols such as resveratrol; chlorogenic acid; catechins; and various proteins and peptides such as casein, whey, casein hydrolysate, whey hydrolysate, MPC (milk protein concentrate), and the tetrapeptide HIRL. The amounts of these ingredients and compounds are within known ranges for which their efficacy has been confirmed.

[0037] The oral composition can be suitably used to enhance CNTF expression in vivo, preferably in the hippocampus, or to promote CNTF production or to improve cognitive function. As described above, enhancing CNTF expression, particularly in the hippocampus, can be expected to suppress the decline in CNTF expression and maintain the survival and promote the differentiation of hippocampal neurons. Furthermore, although not limited to this, the effects of the above or the effects of oleic acid amide and / or palmitic acid amide can also be expected to improve cognitive function. In particular, as shown in the experimental examples described below, ingestion of the oral composition has been confirmed to improve long-term episodic memory. Therefore, the composition is expected to be useful for preventing or ameliorating diseases or symptoms caused by cognitive decline, such as dementia (vascular dementia, Alzheimer's disease, etc.). It is also expected to be useful for preventing or ameliorating amnesia or forgetfulness, even if the disease does not progress to such a condition. Amnesia refers to a decline in memory and is a type of memory disorder. Furthermore, the action of oleic acid amide and / or palmitic acid amide contained in this oral composition is expected to have the effect of suppressing the accumulation of amyloid and tau protein, as well as the effect of enhancing the phagocytic activity and / or anti-inflammatory activity of microglia.

[0038] Therefore, the oral composition can be preferably applied to subjects with cognitive decline or diseases or symptoms resulting from cognitive decline. The cognitive decline includes memory impairment. The cognitive decline also includes age-related cognitive decline, i.e., cognitive decline in healthy middle-aged and elderly individuals and elderly animals. It also includes cognitive decline caused by high-fat diet intake in healthy middle-aged and elderly individuals and elderly animals. The cognitive decline also includes a decline in memory accuracy, spatial awareness, and the ability to understand location.

[0039] As described above, in this specification, the terms "comprise" and "contain" encompass the meanings of "consist of" and "consist essentially of." [Example]

[0040] The present invention will be explained below using experimental examples to aid in understanding the configuration and effects of the present invention. However, the present invention is not limited by these experimental examples. Unless otherwise specified, the following experiments were carried out at room temperature (25±5°C) and atmospheric pressure. Unless otherwise specified, "%" and "parts" in the following descriptions mean "% by mass" and "parts by mass," respectively.

[0041] Manufacturing example: Preparation of Camembert cheese The Camembert cheese used in the experimental examples described below was retort-sterilized Camembert cheese prepared by the following method. Cream with a fat content of 20% was homogenized at 7 MPa (first stage: 5 PMa, second stage: 2 MPa). This homogenized cream was mixed with skim milk with a fat content of 0.1% to prepare raw milk with a fat content of 5.7%. Calcium chloride was added to this raw milk at a rate of 0.025%, and Camembert cheese was produced by conventional methods, including coagulation by adding rennet, cutting and stirring the curd, molding, and draining. The fat content of the solids (FDM) of this Camembert cheese was 62%. This Camembert cheese was heat-treated at a core temperature of 80°C for 10 minutes to prepare heat-treated Camembert cheese. In the following experimental examples, unless otherwise specified, this heat-treated Camembert cheese was used as the "Camembert cheese."

[0042] Experimental Example 1 Ten-week-old male ddY mice were divided into three groups (8 mice per group). Group 1 (Cont: control group) was fed a high-fat diet (5.24 kcal / g: 60% fat, 20% protein, 20% carbohydrate) (D12492: Research Diets, New Brunswick, NJ). Groups 2 (LC: low-dose Camembert cheese group) and 3 (HC: high-dose Camembert cheese group) were fed the high-fat diet plus 8g / kg / day and 15g / kg / day of Camembert cheese, respectively, for one week.

[0043] Group 1 (Cont): High-fat diet Group 2 (LC): High-fat diet + low-dose Camembert cheese (8g / kg / day) Group 3 (HC): High-fat diet + high-dose Camembert cheese (15g / kg / day)

[0044] On the seventh day after the start of feeding, a behavioral test known as a cognitive function evaluation system (episodic memory evaluation test: Object Recognition Test (ORT)) was performed, and on the seventh day, the hippocampus was excised and the expression level of ciliary neurotrophic factor (CNTF) mRNA in the hippocampus was measured.

[0045] (1) Ciliary neurotrophic factor (CNTF) mRNA expression level measurement test (1-1) Test method On day 7 of feeding, mice (n = 8) from each group were anesthetized and dissected, and the hippocampi were removed. The removed hippocampi were stored in RNA later until RNA extraction. RNA was extracted from the hippocampi according to standard methods, and cDNA was synthesized. Subsequently, ciliary neurotrophic factor (CNTF) mRNA expression levels in the hippocampus were measured using RT-PCR according to standard methods. CNTF mRNA expression levels in the hippocampus were also normalized to β-actin mRNA expression levels in the hippocampus.

[0046] (1-2) Test results The test results are shown in Figure 1. Figure 1 shows the relative ratio when the CNTF mRNA expression level in the hippocampus of the high-fat diet group (Group 1: Cont) was set at 1. As shown in Figure 1, compared to the high-fat diet group (Group 1: Cont), the CNTF mRNA expression level in the hippocampus of the high-fat diet + Camembert cheese groups (Group 2: LC, Group 3: HC) increased depending on the amount of Camembert cheese consumed. This revealed that consuming heat-treated Camembert and other white mold cheeses increases the expression of CNTF, which is involved in maintaining the survival and promoting differentiation of nerve cells, particularly hippocampal neurons, and promotes CNTF production, compared to not consuming them.

[0047] (2) Episodic memory evaluation test: Object Recognition Test (ORT): See Figure 2 (2-1) Test method The ORT is a behavioral test that assesses episodic memory, a part of cognitive function. As shown in Figure 2(A), the test consists of three trials: a habituation trial (habituation), an acquisition trial (Trial 1), and a test trial (Trial 2). First, mice were placed in a square open field for 5 min to allow them to habituate to the square open field. After returning them to their home cage, 24 hours later, an acquisition trial (Trial 1) was conducted. In the acquisition trial, two identical objects were placed in the square open field and the mice were allowed to explore freely. When the total exploration time for the two objects reached 20 s, the mice were removed from the square open field, returned to their home cage, and then, 1 hour later, a test trial (Trial 2) was conducted. In the test trial, one of the two objects placed in the square open field was replaced with a novel object of a different shape, and the mice were again placed in the square open field. The mouse's approach time to each object was measured during the test trial. The ORT is an experimental system for assessing cognitive function. If mice remember the shape of the object in the acquisition trial, they will take longer to approach a novel object in the test trial. The cognitive function of mice was assessed using the ratio of the approach time to the novel object to the total approach time to both objects in the test trial. A habituation trial was conducted on day 6 after the start of feeding, and an acquisition trial (Trial 1) and a test trial (Trial 2) were conducted on day 7.

[0048] (2-2) Test results The test results are shown in Figure 2(B). As shown in Figure 2(B), the results of the ORT to evaluate episodic memory revealed that the rate of approaching a novel object was higher in the high-fat diet + Camembert cheese administration groups (LC group, HC group), who also ingested Camembert cheese daily while consuming a high-fat diet, compared to the high-fat diet group (Cont). Furthermore, cognitive function, as assessed by the same ORT, is known to be lower in the high-fat diet group compared to the normal diet group (cognitive decline due to high-fat diet loading). Based on this, it is thought that oral ingestion of heat-treated Camembert and other white mold cheeses may improve cognitive function, particularly episodic memory, which has been impaired by a high-fat diet. In other words, heat-treated white mold cheese has the effect of suppressing cognitive decline and improving long-term memory, especially episodic memory.

[0049] Regarding the Camembert cheese dose of 15g / kg / day for the HC group, the mouse-to-human equivalent dose (HED) conversion factor is 12.3 (according to the US FDA guidelines), so when converted to a human dose it becomes 60g / body (assuming a human body weight of 50kg).

[0050] As shown by the above experimental results, it was confirmed that oral intake of heat-treated white mold cheese (Camembert cheese) increases CNTF expression in the hippocampus, promoting CNTF production, and improving the decline in hippocampal-dependent memory caused by a high-fat diet.

[0051] Experimental Example 2 Experimental Example 1 suggests that heat-treating white mold cheese enhances the effects of suppressing cognitive decline and improving episodic memory. To verify this, the contents of oleic acid amide and palmitic acid amide, which are known to suppress the accumulation of amyloid beta and tau protein associated with brain aging, in white mold cheese were compared between heat-treated and unheated cheeses.

[0052] <Preparation of test cheese> As the white mold cheese used as the test cheese, Camembert cheese prepared by the method described in the above manufacturing example was used. Specifically, as the test cheese of the present invention, six Camembert cheeses (heat-treated cheeses) were prepared according to the description of the above manufacturing example. Further, as the test cheese for comparison, six non-heat-treated Camembert (non-heat-treated cheeses) were prepared in the same manner as the method described in the above manufacturing example except for not performing heat treatment.

[0053] <Method for preparing measurement sample> The amounts of oleic acid amide and palmitic acid amide contained in each of the above test cheeses were measured by LC-MS / MS. Samples for LC-MS / MS were prepared by the following method. [[ID=​​​​​​​​​​​​​​​​​​​​​Column: TOSOH TSKgel, 100v, 2.1 x 75mm, ID 3μm (Part No.21811, Column No.P0014) Column temperature: 40℃ Mobile phase A: 0.1% acetic acid and 5 mM ammonium acetate in water Mobile phase B: 0.1% acetic acid and 5 mM ammonium acetate in water / IPA / acetonitrile (2:58:40) Flow rate: 0.2mL / min Injection volume: 10μL

[0055] [Table 1]

[0056] MS conditions Positive mode [Table 2]

[0057] result As a result of the measurement, the oleic acid amide content of the test cheeses of the present invention (heat-treated cheeses) (n=6) was a minimum of 0.15 μg / g, a maximum of 12.8 μg / g, and an average of 0.55 μg / g. On the other hand, the oleic acid amide content of the test cheeses (unheat-treated cheeses) (n=6) used as a control sample was a minimum of 0.025 μg / g, a maximum of 0.14 μg / g, and an average of 0.08 μg / g. Furthermore, the palmitic acid amide content of the test cheeses of the present invention (heat-treated cheeses) (n=6) was a minimum of 0.18 μg / g, a maximum of 0.45 μg / g, and an average of 0.29 μg / g. On the other hand, the palmitic acid amide content of the test cheeses (unheat-treated cheeses) (n=6) used as control samples was a minimum of 0.02 μg / g, a maximum of 0.1 μg / g, and an average of 0.07 μg / g. These results confirmed that the oleic acid amide and palmitic acid amide contents in white mold cheeses are increased by heat treatment.

[0058] Experimental Example 3 The influence of temperature conditions during heat treatment of white mold cheese on the fatty acid amide content was investigated using oleic acid amide as an example. Camembert cheese (heat-treated cheese) was produced in the same manner as in Experimental Example 2, except for the heating temperature conditions for Camembert cheese, and the oleic acid amide content was measured. The heating temperature conditions used were core temperatures of 70°C, 95°C, and 120°C, and holding times of 30 minutes, 60 minutes, and 120 minutes. However, the holding time at a heating temperature of 120°C was only 30 minutes.

[0059] The results are shown in Figure 3. These results confirmed that the oleamide content in white mold cheese increases with increasing heating time at the same heating temperature, and with increasing heating temperature at the same heating time. In particular, it was confirmed that the oleamide content in white mold cheese increases dramatically with heat treatment equivalent to 95°C for 60 minutes or more. Furthermore, it was confirmed that the oleamide content in white mold cheese increases significantly with heat treatment equivalent to 95°C for 120 minutes or more or 120°C for 30 minutes or more. It is thought that the palmitamide content will also show similar behavior.

Claims

1. An oral composition for enhancing ciliary neurotrophic factor expression, comprising white mold cheese or a processed product thereof as an active ingredient.

2. An oral composition for enhancing ciliary neurotrophic factor expression and improving cognitive function, comprising white mold cheese or a processed product thereof as an active ingredient.

3. An oral composition for enhancing ciliary neurotrophic factor expression and improving cognitive function as described in claim 2, wherein the enhanced expression of ciliary neurotrophic factor is enhanced expression of ciliary neurotrophic factor in the hippocampus.

4. The oral composition according to any one of claims 1 to 3, wherein the white mold cheese is Camembert cheese.

5. The oral composition according to any one of claims 1 to 4, wherein the white mold cheese is heat-treated.

6. The oral composition according to any one of claims 1 to 5, which is a food composition or a food additive composition.

7. A method for producing an oral composition for enhancing ciliary neurotrophic factor expression according to any one of claims 1 and 4 to 6, comprising a step of heat-treating white mold cheese.

8. A method for producing an oral composition for enhancing ciliary neurotrophic factor expression and improving cognitive function according to any one of claims 2 to 6, comprising a step of heat-treating white mold cheese.

9. 9. The method according to claim 7 or 8, wherein the heat treatment step is a step of increasing the content of at least one fatty acid amide selected from the group consisting of oleic acid amide and palmitic acid amide in the white mold cheese.

10. The method according to any one of claims 7 to 9, wherein the white mold cheese is Camembert cheese.

11. A method for producing an oral composition for enhancing ciliary neurotrophic factor expression, or an oral composition for enhancing ciliary neurotrophic factor expression and improving cognitive function, comprising: The manufacturing method is characterized by incorporating white mold cheese or a processed product thereof into the raw material composition of the oral composition for enhancing ciliary neurotrophic factor expression or the oral composition for enhancing ciliary neurotrophic factor expression and improving cognitive function.

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