Ammonia metabolism promoter

Abstract

To provide a raw material having an excellent improving effect of ammonia metabolism.SOLUTION: An ammonia metabolism accelerator having hesperidins and ornithine as active ingredients. Further, an endurance improver, an anti-fatigue agent, and a cognitive function improver. A preventive or an improver of Alzheimer disease in which improvement of a cognitive function is improvement of cognitive function reduction associated with excessive accumulation ammonia, and improvement of cognitive function reduction associated with Alzheimer type dementia. Further, a mass ratio between hesperidins and ornithine is 1:0.1 to 10.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a material that promotes ammonia metabolism in the body. [Background technology]

[0002] Ammonia in the body is mainly generated during the process of catabolism of amino acids in protein metabolism or muscle breakdown due to exercise. Ammonia inhibits the oxidation of pyruvate to acetyl CoA, causing fatigue (see Non-Patent Document 1). It is also known that ammonia reduces the function of the nervous system, causing central fatigue, and promotes the accumulation of lactic acid in muscles, promoting muscle fatigue (Patent Document 1).

[0003] Ammonia in the body is converted into non-toxic urea by the urea cycle in the liver and excreted in urine. Arginine, ornithine and citrulline are involved in the urea cycle in the liver, which converts ammonia into non-toxic urea. It has been reported that ornithine, citrulline or arginine, which are amino acids involved in the urea cycle, suppress the increase in blood ammonia (Patent Documents 2 and 3, and Non-Patent Document 1). In the body, citrulline is synthesized from ornithine, and citrulline is converted to arginine. It has been reported that ornithine suppresses the increase in blood ammonia and improves fatigue (see Patent Document 2). It has also been reported that citrulline intake increases blood arginine and improves brain fatigue (Patent Documents 3 and 4).

[0004] Endurance is not only necessary in athletic competitions, but is also essential in our daily lives, for simple movements such as walking and running, and for labor that involves repeated muscular exertion. A decline in endurance, especially in the elderly, can cause difficulties in daily movements and have a significant negative impact on quality of life (QOL). It is widely recognized that exercise training is effective in improving endurance, but maintaining or improving endurance through training is not easy due to the difficulty of finding time and maintaining motivation, and the risk of injury for the elderly.

[0005] As ingredients that have the effect of improving endurance or preventing fatigue, various amino acids have been reported, such as ornithine (Patent Document 2), citrulline (Non-Patent Document 1), a combination of citrulline and arginine (Patent Document 3), a combination of arginine, ornithine and citrulline (Non-Patent Document 2), etc. In addition, a combination of catechins and ornithine has been reported as an ingredient that promotes ammonia metabolism, improves endurance and prevents fatigue (Patent Document 5).

[0006] Ammonia is also a normal end product of human tissue metabolism that has been demonstrated to be toxic to brain function; for example, elevated blood ammonia levels due to impaired liver function reduces brain function, known as hepatic encephalopathy. In addition, various associations between blood ammonia levels and Alzheimer's disease have been proposed and suggested (Non-Patent Documents 3-7), and it has recently been reported that, depending on the duration and magnitude of ammonia exposure, elevated brain ammonia causes typical neurological symptoms common to Alzheimer's disease, including memory dysfunction, cognitive and spatial learning disorders, and pathophysiologically, mitochondrial dysfunction, disruption of cellular glucose metabolism, impaired neurotransmission, and inflammatory responses occur, and it has been reported that the conventional assumption that chronically elevated ammonia levels are associated with the onset of Alzheimer's disease is valid (Non-Patent Document 8).

[0007] In Non-Patent Document 8, the relationship between Alzheimer's disease brains and ammonia is discussed as follows. In the brain, in either normal or hyperammoniac conditions, the pathway of ATP-dependent formation of glutamine by glutamine synthetase (1-glutamate: ammonia ligase (ADP formation; EC6.3.1.2)) (GS) is mainly used for ammonia removal. In Alzheimer's disease brains, the extracellular deposition density of Aβ and senile plaques (SP) in the cortex not only enhances the neurotoxicity of Aβ but also induces oxidative structural changes of GS by interacting with it. This causes a loss of GS activity in astrocytes, inducing impairment of the brain's ammonia clearance mechanism (glutamate-glutamine cycle) and astrocytes. That is, Non-Patent Document 8 shows that the extracellular deposition of Aβ and senile plaques (SP) in the brain of Alzheimer's disease not only causes neurological disorders itself, but also reduces the brain's ammonia clearance ability, causing cognitive decline, and suggests the importance of reducing ammonia in Alzheimer's disease.

[0008] Furthermore, Non-Patent Document 8 describes that a diet containing a large amount of lactic acid bacteria is effective in treating Alzheimer's disease, and the mechanism is considered to be that the lactic acid bacteria suppress ammonia production or absorption from the digestive tract, resulting in a decrease in blood ammonia levels. Therefore, it is believed that reducing blood ammonia levels by acting on the periphery may be effective in treating Alzheimer's disease.

[0009] On the other hand, hesperidins are flavonoids that are abundant in citrus fruits, and have been reported to have physiological effects such as strengthening capillaries, improving blood flow, and anti-allergic effects. However, there have been no reports on the effects of hesperidin on ammonia metabolism. In addition, there have been no reports on the effects of hesperidin in combination with ornithine. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] Japanese Unexamined Patent Application Publication No. 2000-239179 [Patent Document 2] Japanese Unexamined Patent Application Publication No. 2011-132174 [Patent Document 3] International Publication No. 2009 / 048148 [Patent Document 4] Japanese Unexamined Patent Application Publication No. 2013-060406 [Patent Document 5] Japanese Unexamined Patent Application Publication No. 2018-203719 [Non-Patent Document]

[0011] [Non-Patent Document 1] Takeda et al, J Nutr Sci Vitaminol, 2011, 57:246-250 [Non-Patent Document 2] Meneguello et al, Cell Biochem Funct, 2003, 21:85-91 [Non-Patent Document 3] Fisman M. et.al., J. Am. Geriatr. Soc., 1989, 37, 1102-1102 [Non-Patent Document 4] Fisman M. et.al., AM. J. PSYCHIATRY, 1985, 142, 71-73 [Non-Patent Document 5] Hoyer S., ADV. EXP. MED. BIOL., 1994, 368, 197-205 [Non-Patent Document 6] Seiler N., Neurochem. Int., 2002, 41, 189-207 [Non-Patent Document 7] Seiler N., Neurochem Res., 1993, 18, 235-245 [Non-Patent Document 8] Jin Y.Y. et.al., Nutrients, 2018, 10, 564 Summary of the Invention [Problem to be solved by the invention]

[0012] The present invention relates to providing a material having an excellent ammonia metabolism promoting effect. [Means for solving the problem]

[0013] The present inventors have found that the combined use of hesperidins and ornithine significantly improves ammonia metabolism in hepatocytes, as compared with the use of hesperidins or ornithine alone.

[0014] Therefore, the present invention provides an ammonia metabolism promoter containing hesperidins and ornithine as active ingredients. The present invention also provides an endurance improver containing hesperidins and ornithine as active ingredients. The present invention also provides an anti-fatigue agent containing hesperidins and ornithine as active ingredients. The present invention also provides an agent for improving cognitive function, which comprises hesperidins and ornithine as active ingredients. The present invention also provides an agent for preventing or ameliorating Alzheimer's disease, which comprises hesperidins and ornithine as active ingredients. The present invention further provides a food for promoting ammonia metabolism, which contains hesperidins and ornithine as active ingredients. The present invention also provides a food for improving endurance, which contains hesperidins and ornithine as active ingredients. The present invention also provides an anti-fatigue food containing hesperidins and ornithine as active ingredients. The present invention also provides a food for improving cognitive function, which contains hesperidins and ornithine as active ingredients. The present invention also provides a food for preventing or ameliorating Alzheimer's disease, which contains hesperidins and ornithine as active ingredients. Effect of the Invention

[0015] According to the present invention, ammonia metabolism in the body can be promoted, and it becomes possible to improve endurance, relieve fatigue, improve cognitive function, and prevent or ameliorate disorders accompanied by decline in cognitive function, such as Alzheimer's disease. [Brief description of the drawings]

[0016] [Figure 1] Effects of hesperidin and ornithine on ammonia metabolism (urea production) in human iPS cell-derived hepatocytes. The horizontal axis represents the human hepatocyte culture medium, and the vertical axis represents the urea concentration in the medium. Data are the mean ± SE (n = 4), *p < 0.05 (Dunnett test, vs. control group). [Diagram 2] Effects of hesperidins and ornithine on ammonia metabolism (urea production) in human iPS cell-derived hepatocytes. The horizontal axis represents the human hepatocyte culture medium, and the vertical axis represents the urea concentration in the medium. Data are the mean ± SE (n = 4), *p < 0.05 (Dunnett test, vs. control group). [Diagram 3] The effect of urea cycle activators on ammonia metabolism (urea production) in human iPS cell-derived hepatocytes. The horizontal axis represents the human hepatocyte culture medium, and the vertical axis represents the urea concentration in the medium. Data are mean ± SE (n = 4), *p < 0.05 (t-test). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] All patents, non-patent documents, and other publications cited herein are hereby incorporated by reference in their entirety.

[0018] As used herein, "non-therapeutic" is a concept that does not include medical procedures, i.e., methods of surgery, treatment, or diagnosis of humans, and more specifically, does not include methods of surgery, treatment, or diagnosis performed on humans by a physician, medical professional, or a person under the direction of a physician.

[0019] As used herein, "prevention" refers to preventing, suppressing, or delaying the onset of a disease or condition in an individual, or reducing an individual's risk of developing a disease or condition. Also, as used herein, "amelioration" refers to improving a disease or condition, preventing, suppressing, or delaying the worsening of a disease or condition, or reversing, preventing, suppressing, or delaying the progression of a disease or condition.

[0020] As used herein, "exercise" refers to physical exercise in a broad sense, including, for example, athletic competitions such as sports, training, and aerobic exercise (exercise in the narrow sense), as well as work involving muscular exertion, daily movements, etc. As used herein, when "exercise" is used in relation to athletes or those who exercise in the narrow sense, it may preferably mean exercise in the narrow sense, whereas when "exercise" is used in relation to middle-aged or elderly people, the infirm, sick, or those recovering from illness, it may preferably mean work involving muscular exertion or daily movements.

[0021] In this specification, "endurance" refers to endurance for the above-mentioned "exercise," and "improving endurance" is a concept that includes increasing endurance and suppressing a decrease in endurance. Furthermore, in this specification, "anti-fatigue" refers to suppressing fatigue caused by the above-mentioned "exercise" or promoting recovery from it. Suppressing fatigue caused by exercise or promoting recovery from it leads to improved endurance.

[0022] As used herein, "cognitive function" refers to memory, learning, thinking, attention or perception, language, spatiotemporal awareness, orientation, and other abstract phenomena recognition or comprehensive judgment ability, and executive ability, and preferably includes memory or learning ability, comprehensive judgment ability, and executive ability. "Improvement of cognitive function" means the maintenance and improvement of cognitive function, and the alleviation and cure of various symptoms (cognitive dysfunction) associated with the decline of cognitive function. Examples of diseases or conditions that present with cognitive dysfunction include dementia (e.g., senile dementia, Alzheimer's disease, vascular dementia, post-traumatic dementia, as well as dementia caused by various diseases such as brain tumor, chronic subdural hematoma, normal pressure cerebral edema, meningitis, Parkinson's disease, etc.), non-dementia cognitive impairment (e.g., mild cognitive impairment (MCI)), memory or learning disorder (e.g., memory and learning disorder associated with brain development disorder), etc., and preferably Alzheimer's disease or Alzheimer's dementia.

[0023] As used herein, "hesperidins" means at least one selected from the group consisting of hesperidin, methylated hesperidin (methylhesperidin), and sugar adducts in which sugar is bound to hesperidin (hesperidin sugar adducts). Hesperidin is a glycoside in which rutinose (L-rhamnosyl-(α1→6)-D-glucose) is beta-linked to the 7-hydroxyl group of hesperetin (5,7,3'-trihydroxy-4'-methoxyflavanone). Hesperidin can be industrially produced by known methods utilizing chemical synthesis or enzymatic reactions, or can be obtained by extraction from natural products containing hesperidin, preferably plants. As the hesperidin, commercially available hesperidin-containing preparations may be used, for example, "Hesperidin" (Hamari Pharmaceutical Co., Ltd.) and "Hesperidin" (Alps Pharmaceutical Co., Ltd.).

[0024] The position and number of methylations in methylhesperidin are not particularly limited. Methylhesperidin is known to mainly contain chalcone type compounds (1) and flavanone type compounds (2), and examples of its components include those having the structures shown below.

[0025] [ka]

[0026] (wherein R represents a hydrogen atom or a methyl group).

[0027] Here, methylhesperidin as a pharmaceutical additive and food additive is mainly handled as a mixture of compounds (3) and (4).

[0028] [ka]

[0029] (In the formula, Gl represents a glucose residue, and Rh represents a rhamnose residue. Gl-2 represents the 2-position of the glucose residue (including the 3-position in the case of (3-1)), and Rh-2 represents the 2-position of the rhamnose residue.)

[0030] In the present invention, methylhesperidin may contain both the chalcone type compound (1) and the flavanone type compound (2) shown above, or may contain only one of them. In the present invention, a more preferred methylhesperidin is a mixture of compound (3) and compound (4).

[0031] Methylhesperidin can be produced by known methods, for example, by dissolving hesperidin in an aqueous sodium hydroxide solution, reacting the alkaline solution with a corresponding amount of dimethyl sulfate, neutralizing the reaction solution with sulfuric acid, extracting with n-butyl alcohol, distilling off the solvent, and then recrystallizing with isopropyl alcohol (Sakiyoku, Nippon Kagaku Zasshi, 79, 733-6 (1958)), but the production method is not limited to this. As methylhesperidin, commercially available methylhesperidin-containing preparations may be used, such as "methylhesperidin" (Tokyo Chemical Industry Co., Ltd.), "hesperidin methylchalcone" (Sigma), "methylhesperidin" (Hamari Pharmaceutical Co., Ltd.), "methylhesperidin" (Showa Denko K.K.), and "methylhesperidin" (Alps Pharmaceutical Co., Ltd.).

[0032] A hesperidin sugar adduct is a compound in which another sugar, such as glucose, maltose, fructose, rhamnose, lactose, etc., is bound to the sugar portion (rutinose portion) of hesperidin. In the present invention, the sugar adduct of hesperidin is preferably glucosylhesperidin in which 1 to 10 glucose molecules are bound to hesperidin, and more preferably monoglucosylhesperidin in which one glucose molecule is bound, from the viewpoint of solubility. The number of glucose molecules added may vary, and the average number of moles of glucose added per mole of hesperidin is preferably 1 to 10.

[0033] Hesperidin sugar adducts can be produced by known methods, for example, by reacting hesperidin with a glycosyltransferase in the presence of a sugar donor (see the description in the specification of Japanese Patent No. 3060227, etc.), but the production method is not limited to this. As the glucosyl hesperidin, commercially available glucosyl hesperidin-containing preparations may be used, such as monoglucosyl hesperidin ("αG Hesperidin PA-T" (Toyo Sugar Refining Co., Ltd.) and "Hayashibara Hesperidin (registered trademark) S" (Hayashibara Co., Ltd.).

[0034] In the present invention, ornithine can be used in the form of a free form or a salt thereof. Ornithine may be L-form, D-form, DL-form, or a mixture thereof, but is preferably L-form.

[0035] Examples of the salt of ornithine include acid addition salts, metal salts, ammonium salts, organic amine addition salts, and amino acid addition salts. Examples of the acid addition salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, and phosphate, and organic acid salts such as acetate, maleate, fumarate, citrate, malate, lactate, α-ketoglutarate, gluconate, and caprylate. Examples of the metal salts include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt, and zinc salt. Examples of the ammonium salts include salts such as ammonium and tetramethylammonium. Examples of the organic amine addition salts include salts such as morpholine and piperidine. Examples of the amino acid addition salts include salts of glycine, phenylalanine, lysine, aspartic acid, and glutamic acid. Among these, sodium salt or hydrochloride is preferred.

[0036] The free ornithine or a salt thereof can be obtained by a method of isolating and purifying from animals or plants containing them, chemical synthesis, fermentation production, etc. Alternatively, a commercially available product may be purchased.

[0037] In the present invention, by using hesperidins and ornithine in combination, a significantly higher ammonia metabolism promoting effect is achieved compared to the application of hesperidins or ornithine alone. As mentioned above, ammonia inhibits the oxidation of pyruvate to acetyl CoA, causing fatigue (Non-Patent Document 1). In addition, suppressing fatigue caused by exercise or promoting its recovery leads to improved endurance. Furthermore, as mentioned above, high concentrations of ammonia accumulate in the blood, and a portion of it migrates into the brain, causing various neurological symptoms. For example, it has been reported that depending on the duration and magnitude of ammonia exposure, an increase in brain ammonia causes typical neurological symptoms common to Alzheimer's disease, including memory dysfunction, cognitive and spatial learning disorders, and that reducing blood ammonia concentration is effective in treating Alzheimer's disease (Non-Patent Document 8).

[0038] Therefore, the combination of hesperidins and ornithine, which have excellent ammonia metabolism promoting effect, can be an ammonia metabolism promoter, an endurance improver, an anti-fatigue agent, a cognitive function improver (preferably, an improver for cognitive function decline accompanied by excessive accumulation of ammonia, an improver for cognitive function decline caused by Alzheimer's dementia), and even an agent for preventing or improving Alzheimer's disease.In addition, in the present invention, the combination of hesperidins and ornithine can be administered simultaneously, or each component can be administered separately, as long as they can cooperate in vivo.

[0039] In one aspect, the present invention provides an ammonia metabolism promoter containing hesperidins and ornithine as active ingredients.The present invention also provides an endurance improver containing hesperidins and ornithine as active ingredients.The present invention also provides an anti-fatigue agent containing hesperidins and ornithine as active ingredients.The present invention also provides a cognitive function improver containing hesperidins and ornithine as active ingredients.The present invention also provides an agent for preventing or improving Alzheimer's disease containing hesperidins and ornithine as active ingredients.

[0040] In another aspect, the present invention provides use of hesperidins and ornithine for the manufacture of an agent for promoting ammonia metabolism, an agent for improving endurance, an anti-fatigue agent, an agent for improving cognitive function, or an agent for the prevention or amelioration of Alzheimer's disease.

[0041] In one embodiment, the ammonia metabolism enhancer, endurance enhancer, anti-fatigue agent, cognitive function improver, and Alzheimer's disease preventive or ameliorating agent of the present invention is a composition containing at least hesperidins and ornithine. In another embodiment, the agent of the present invention may essentially consist of hesperidins and ornithine.

[0042] In another aspect, the present invention provides use of hesperidins and ornithine for promoting ammonia metabolism, improving stamina, preventing fatigue, improving cognitive function, or preventing or improving Alzheimer's disease. In a preferred embodiment, the hesperidins and ornithine are used in the form of a composition containing them.

[0043] In yet another aspect, the present invention provides a combination of hesperidins and ornithine for use in promoting ammonia metabolism, improving stamina, fighting fatigue, improving cognitive function, or preventing or improving Alzheimer's disease.In a preferred embodiment, the combination of hesperidins and ornithine is a composition containing them.

[0044] Examples of compositions containing hesperidins and ornithine include medicines, quasi-drugs, and foods, which will be described later.

[0045] The use of hesperidins and ornithine according to the present invention can include therapeutic use and non-therapeutic use.Non-therapeutic use can include, for example, administering or ingesting hesperidins and ornithine for the purpose of obtaining health-promoting effects such as reducing fatigue caused by physical activity in daily life, improving endurance and reducing fatigue in labor involving muscular labor, improving stamina during exercise in the narrow sense, promoting recovery from fatigue during or after exercise in the narrow sense, and maintaining or improving cognitive function, without being a medical procedure.In addition, for example, non-therapeutic use can include providing hesperidins and ornithine to others, claiming the purpose of the above-mentioned health-promoting effects, so that they can be administered or ingested by others without being a medical procedure.

[0046] Therapeutic uses include applications in the prevention or amelioration of diseases or conditions caused by excessive accumulation of ammonia, such as chronic fatigue syndrome, hyperammonemia, and hepatic encephalopathy, or in the prevention or amelioration of diseases or conditions presenting with cognitive dysfunction, such as Alzheimer's disease.

[0047] In the present invention, hesperidins and ornithine can be used for both human and non-human animals, but are preferably used for human.Non-human animals include non-human mammals, amphibians, and cartilaginous fish, and non-human mammals include apes, other primates, mice, rats, horses, cows, pigs, sheep, dogs, cats, hamsters, and companion animals.

[0048] In yet another aspect, the present invention provides a method for promoting ammonia metabolism in a subject. The present invention also provides a method for improving endurance in a subject. The present invention also provides a method for anti-fatigue in a subject. The present invention also provides a method for improving cognitive function in a subject. The present invention also provides a method for preventing or ameliorating Alzheimer's disease in a subject. The method comprises administering to the subject an effective amount of hesperidins and ornithine. In a preferred embodiment, a composition comprising hesperidins and ornithine is administered. In another preferred embodiment, the administration is oral administration.

[0049] Examples of subjects to which the method of the present invention is applied include the above-mentioned humans and non-human animals in need of promotion of ammonia metabolism, improvement of stamina, anti-fatigue, or improvement of cognitive function, as well as the above-mentioned humans and non-human animals in need of prevention or amelioration of diseases or conditions caused by excessive accumulation of ammonia, such as chronic fatigue syndrome, hyperammonemia, or hepatic encephalopathy, or prevention or amelioration of diseases or conditions presenting with cognitive dysfunction, such as Alzheimer's disease. Further examples of such subjects include the above-mentioned humans and non-human animals in need of reduction in fatigue due to physical activity in daily life, improvement of durability and reduction of fatigue in work involving muscular exertion, improvement of endurance during exercise in the narrow sense, promotion of recovery from fatigue during or after exercise in the narrow sense, maintenance and improvement of cognitive function, etc. Further examples of such subjects include conditions caused by hyperammonemia, such as respiratory alkalosis, slurred speech, tremors, weakness, increased or decreased muscle tone, ataxia, hypothermia, seizures, brain oedema, coma, brain stem compression, confusion with headache, nausea, vomiting, agitation, delusions and delirium, irritability, aggression, hyperactivity, hyperactive, bizarre or self-injurious behavior, cognitive deficits, protein aversion, The present invention also includes the above-mentioned humans and non-human animals in need of prevention or improvement of disorders such as aversion, anorexia, delayed growth, and the like. Further examples of such subjects include the above-mentioned humans and non-human animals in need of suppression of central fatigue associated with decreased function of the nervous system due to ammonia accumulation, or suppression of sleepiness or fatigue due to said central fatigue. Further examples of such subjects include the humans and non-human animals described above in need of prevention or amelioration of symptoms or conditions associated with elevated blood ammonia levels, such as slurred speech, lethargy, coma, impaired consciousness, blurred vision, developmental disorders, protein-induced vomiting, and impaired feeding in infants. Further examples of such subjects include the above-mentioned humans and non-human animals in need of prevention or improvement of diseases or conditions presenting with cognitive impairment, such as dementia (e.g., senile dementia, Alzheimer's disease, vascular dementia, post-traumatic dementia, as well as dementia caused by various diseases such as brain tumor, chronic subdural hematoma, normal pressure cerebral edema, meningitis, Parkinson's disease, etc.), non-dementia cognitive impairment (e.g., mild cognitive impairment (MCI)), memory or learning disorder (e.g., memory and learning disorder associated with brain development disorder), etc. Alternatively, the method for promoting ammonia metabolism of the present invention may be an in vitro method, and subjects to which the in vitro method is applied include liver tissue derived from the above-mentioned human or non-human animals, cultured hepatocytes (e.g., primary cultured liver cells), iPS cell-derived hepatocytes, and the like.

[0050] The effective amount of administration in the method of the present invention may be an amount capable of achieving the promotion of ammonia metabolism in a subject, or an amount capable of achieving the improvement of endurance in a subject, or an amount capable of achieving anti-fatigue in a subject, or an amount capable of achieving the improvement of cognitive function in a subject, or an amount capable of preventing or improving Alzheimer's disease in a subject. The ammonia metabolism level can be evaluated by measuring the blood ammonia concentration, urea production amount, ammonia concentration in a culture medium of liver cells, urea concentration, etc. of a subject. Endurance and fatigue resistance can be evaluated by exercise tests such as a treadmill test and a bicycle pedaling test. Cognitive function can be evaluated by neuropsychological tests and brain imaging tests. In one embodiment, the effective amount may be an amount that statistically significantly increases the ammonia metabolism level of a group administered with hesperidins and ornithine compared to a non-administered group. In another embodiment, the effective amount may be an amount that statistically significantly reduces the blood ammonia concentration of a group administered with hesperidins and ornithine compared to a non-administered group. In yet another embodiment, the effective amount can be the amount that statistically significantly extends the exercise time of the group of individuals administered with hesperidins and ornithine compared to the group that is not administered with hesperidins and ornithine.In yet another embodiment, the effective amount can be the amount that statistically significantly improves the cognitive function of the group of individuals administered with hesperidins and ornithine compared to the group that is not administered with hesperidins and ornithine.

[0051] In the examples described later, the ammonia metabolism level is evaluated by measuring the urea concentration in the culture medium of human iPS cell-derived hepatocytes. Human iPS cell-derived hepatocytes exhibit characteristics very similar to those of human primary culture hepatocytes in terms of morphology, expressed proteins, metabolic activity, etc., and homogeneous hepatocytes can be obtained stably and in large quantities, and biological test results are highly reproducible. As shown in the examples described later, when a urea cycle activator was applied to human iPS cell-derived hepatocytes, the urea concentration in the culture medium significantly increased. This revealed that the urea cycle functions normally in human iPS cell-derived hepatocytes, and demonstrated that a test system using human iPS cell-derived hepatocytes is preferably used to evaluate the ammonia metabolism level. Human iPS cell-derived hepatocytes can be produced by a known method. Alternatively, commercially available products may be purchased, and examples of such commercially available products include iCell (registered trademark) Hepatocytes 2.0 01279 strain (FUJIFILM Wako Pure Chemical Industries, Ltd.) and ReproHepato (trademark) (ReproCell Co., Ltd.).

[0052] In the present invention, hesperidins and ornithine can be used as active ingredients for imparting the functions of promoting ammonia metabolism, improving stamina, anti-fatigue, or improving cognitive function to medicines, quasi-drugs, or foods (including foods for non-human animals), or as active ingredients for preventing or improving diseases or conditions caused by excessive accumulation of ammonia, such as chronic fatigue syndrome, hyperammonemia, and hepatic encephalopathy, or for preventing or improving diseases or conditions presenting with cognitive dysfunction, such as Alzheimer's disease.

[0053] The pharmaceutical (including quasi-drugs) is a pharmaceutical for promoting ammonia metabolism, improving stamina, anti-fatigue, or improving cognitive function, or for preventing or improving diseases or conditions caused by excessive accumulation of ammonia, such as chronic fatigue syndrome, hyperammonemia, and hepatic encephalopathy, or for preventing or improving diseases or conditions that present cognitive dysfunction, such as Alzheimer's disease, and contains hesperidins and ornithine as active ingredients for the functions. Furthermore, the pharmaceutical may contain a pharmaceutical carrier, other active ingredients, pharmacological ingredients, etc., as necessary, as long as the function of the active ingredient is not lost. The pharmaceutical may be provided as one composition containing hesperidins and ornithine, or as a combination of multiple compositions containing hesperidins or ornithine.

[0054] The dosage form of this medicine (including quasi-drug) can be either oral administration or parenteral administration, but oral administration is preferred.The dosage form of this medicine is not particularly limited as long as it is oral or parenterally administrable, and can be, for example, injection, suppository, inhalant, transdermal absorption agent, various external preparations, tablet, capsule, granule, powder, liquid, syrup, etc., and the preparation of such various dosage forms can be appropriately combined with hesperidins and ornithine and pharmacologic acceptable carriers (for example, excipients, binders, bulking agents, disintegrants, surfactants, lubricants, dispersants, buffers, preservatives, flavorings, flavorings, coating agents, diluents, etc.), other medicinal ingredients, etc., and prepared according to standard methods.

[0055] The food is a food for providing the functions of promoting ammonia metabolism, improving stamina, anti-fatigue, or improving cognitive function, and contains hesperidins and ornithine as active ingredients for the functions. The food includes foods for sick people based on the concept of promoting ammonia metabolism, improving stamina, anti-fatigue, or improving cognitive function, and labeled as such as necessary, as well as health functional foods such as nutritional functional foods, foods for specified health uses, and foods with functional claims.

[0056] The food provided by the present invention includes beverages. Therefore, the food can be referred to as food and drink. The form of the food can be solid, semi-solid or liquid (e.g. beverages). Examples of the food include bread, noodles, rice, cookies and other confectionery, jellies, dairy products, soups, frozen foods, instant foods, starch processed products, processed fish products, other processed foods, seasonings, nutritional supplements, and beverages such as tea and coffee drinks, fruit drinks, carbonated drinks, and jelly-like drinks, as well as their raw materials. Alternatively, the food may be a supplement having the form of an oral administration preparation such as a tablet, capsule, granule, powder, liquid, or syrup. The food may be provided as one composition containing hesperidins and ornithine, or as a combination of multiple compositions containing hesperidins or ornithine.

[0057] The food can be prepared according to standard methods by appropriately combining hesperidins and ornithine with any food ingredients or additives acceptable for food (e.g., solvents, softeners, oils, emulsifiers, preservatives, flavorings, sweeteners, stabilizers, colorants, UV absorbers, antioxidants, moisturizers, thickeners, adhesives, dispersants, humectants, etc.).

[0058] The content of hesperidins in the pharmaceutical product (including quasi-drugs) is not particularly limited, but is preferably 0.1% by mass or more, more preferably 1% by mass or more, and even more preferably 3% by mass or more, and is preferably 20% by mass or less, more preferably 10% by mass or less, and even more preferably 5% by mass or less, calculated as hesperidin. Examples of the content include 0.1 to 20% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 1 to 20% by mass, 1 to 10% by mass, 1 to 5% by mass, 3 to 20% by mass, 3 to 10% by mass, and 3 to 5% by mass.

[0059] The content of ornithine in the pharmaceutical (including quasi-drug) is not particularly limited, but is preferably 0.3% by mass or more, more preferably 3% by mass or more, even more preferably 5% by mass or more, even more preferably 9% by mass or more, and is preferably 60% by mass or less, more preferably 30% by mass or less, even more preferably 20% by mass or less, even more preferably 15% by mass or less, in terms of free ornithine. Further, examples of the content include 0.3 to 60% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 3 to 60% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 5 to 60% by mass, 5 to 30% by mass, 5 to 20% by mass, 5 to 15% by mass, 9 to 60% by mass, 9 to 30% by mass, 9 to 20% by mass, and 9 to 15% by mass.

[0060] In the pharmaceutical product (including quasi-drugs), any of the above-listed hesperidin concentrations (0.1 to 20 mass%, 0.1 to 10 mass%, 0.1 to 5 mass%, 1 to 20 mass%, 1 to 10 mass%, 1 to 5 mass%, 3 to 20 mass%, 3 to 10 mass%, and 3 to 5 mass%) can be arbitrarily combined with any of the above-listed ornithine concentrations (0.3 to 60 mass%, 0.3 to 30 mass%, 0.3 to 20 mass%, 0.3 to 15 mass%, 3 to 60 mass%, 3 to 30 mass%, 3 to 20 mass%, 3 to 15 mass%, 5 to 60 mass%, 5 to 30 mass%, 5 to 20 mass%, 5 to 15 mass%, 9 to 60 mass%, 9 to 30 mass%, 9 to 20 mass%, and 9 to 15 mass%).

[0061] The content of hesperidins in the food is not particularly limited, but is preferably 0.05% by mass or more, more preferably 0.1% by mass or more, and even more preferably 1% by mass or more, and is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 1% by mass or less, in terms of hesperidin. Examples of the content include 0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, and 1 to 5% by mass.

[0062] The content of ornithine in the food is not particularly limited, but is preferably 0.15% by mass or more, more preferably 0.3% by mass or more, more preferably 3% by mass or more, more preferably 5% by mass or more, and is preferably 30% by mass or less, more preferably 20% by mass or less, more preferably 15% by mass or less, more preferably 5% by mass or less, and more preferably 3% by mass or less. Further, examples of the content include 0.15-30% by mass, 0.15-20% by mass, 0.15-15% by mass, 0.15-5% by mass, 0.15-3% by mass, 0.3-30% by mass, 0.3-20% by mass, 0.3-15% by mass, 0.3-5% by mass, 0.3-3% by mass, 3-30% by mass, 3-20% by mass, 3-15% by mass, 3-5% by mass, 5-30% by mass, 5-20% by mass, and 5-15% by mass.

[0063] In the food, any of the above-listed hesperidin concentrations (0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, and 1 to 5% by mass) can be arbitrarily combined with any of the above-listed ornithine concentrations (0.15 to 30% by mass, 0.15 to 20% by mass, 0.15 to 15% by mass, 0.15 to 5% by mass, 0.15 to 3% by mass, 0.3 to 20% by mass, 0.3 to 30% by mass, 0.3 to 15% by mass, 0.3 to 5% by mass, 0.3 to 3% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 3 to 5% by mass, 5 to 30% by mass, 5 to 20% by mass, and 5 to 15% by mass).

[0064] In the present invention, the dosage and administration schedule of hesperidins and ornithine may be appropriately determined by those skilled in the art according to the species, body weight, sex, age, condition, or other factors of the subject. Without being limited thereto, in the case of oral administration, examples of the dosage of hesperidins and ornithine according to the present invention per day for an adult are as follows: [Hesperidins (converted into hesperidin equivalent)] preferably 100 to 3000 mg / 60 kg body weight, more preferably 250 to 2000 mg / 60 kg body weight, even more preferably 250 to 1000 mg / 60 kg body weight, still more preferably 250 to 600 mg / 60 kg body weight; [Ornithine (free form equivalent)] preferably 100 to 5000 mg / 60 kg body weight, more preferably 250 to 3000 mg / 60 kg body weight, even more preferably 400 to 2000 mg / 60 kg body weight, even more preferably 500 to 2000 mg / 60 kg body weight, even more preferably 800 to 1600 mg / 60 kg body weight, in another example, preferably 250 to 800 mg / 60 kg body weight, even more preferably 500 to 800 mg / 60 kg body weight, The above doses are preferably administered, for example, once a day, or divided into two or three or more doses a day.

[0065] The ratio by mass of hesperidins (converted into hesperidin) to ornithine (converted into free form) used in the present invention is preferably 1:0.1 to 10, more preferably 1:0.5 to 10, even more preferably 1:1 to 8, even more preferably 1:1 to 5, and even more preferably 1:2 to 4. For example, the ratio by mass of hesperidins to ornithine in the ammonia metabolism promoter, endurance improver, anti-fatigue agent, cognitive function improver, and Alzheimer's disease preventive or ameliorating agent of the present invention is preferably 1:0.1 to 10, more preferably 1:0.5 to 10, even more preferably 1:1 to 8, even more preferably 1:1 to 5, and even more preferably 1:2 to 4. Furthermore, for example, the dosage ratio of hesperidins to ornithine administered in the present invention's methods for promoting ammonia metabolism, enhancing endurance, anti-fatigue, or improving cognitive function, the methods for preventing or improving diseases or conditions presenting with cognitive dysfunction such as Alzheimer's disease, and the methods for preventing or improving diseases or conditions caused by excessive accumulation of ammonia is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, even more preferably 1:1-5, and still more preferably 1:2-4, by mass.

[0066] In a preferred embodiment of the non-therapeutic use of hesperidins and ornithine for promoting ammonia metabolism, improving stamina, anti-fatigue, or improving cognitive function according to the present invention, the hesperidins and ornithine are provided as a composition containing 1 to 10% by mass (converted into hesperidin) of hesperidins and 5 to 20% by mass (converted into free form) of ornithine, and the mass ratio of hesperidins (converted into hesperidin) to ornithine (converted into free form) is 1:0.5 to 10. Preferably, the composition is a food product. In a preferred embodiment of the non-therapeutic use of hesperidins and ornithine for promoting ammonia metabolism, improving endurance, anti-fatigue, or improving cognitive function according to the present invention, the hesperidins and ornithine are provided as a beverage composition containing 0.057 to 0.567% by mass (converted into hesperidin) of hesperidins and 0.283 to 1.132% by mass (converted into free form) of ornithine, and in which the mass ratio of hesperidins (converted into hesperidin) to ornithine (converted into free form) is 1:0.5-10. In a preferred embodiment of the therapeutic use of hesperidins and ornithine according to the present invention for promoting ammonia metabolism, enhancing stamina, anti-fatigue, or improving cognitive function, for preventing or ameliorating Alzheimer's disease, or for preventing or ameliorating chronic fatigue syndrome, hyperammonemia, or hepatic encephalopathy, the hesperidins and ornithine are provided as a composition containing 1 to 10% by mass (converted into hesperidin) of hesperidins and 5 to 20% by mass (converted into free form) of ornithine, and the mass ratio of hesperidins (converted into hesperidin) to ornithine (converted into free form) is 1:0.5 to 10. Preferably, the composition is a pharmaceutical product. In the present invention, the preferred dosage of hesperidins and ornithine used for promoting ammonia metabolism, improving stamina, anti-fatigue, or improving cognitive function, preventing or improving Alzheimer's disease, or preventing or improving chronic fatigue syndrome, hyperammonemia, or hepatic encephalopathy is 250-1000mg of hesperidins (converted into hesperidin) / 60kg body weight, and 400-2000mg of ornithine (converted into free form) / 60kg body weight as an oral dosage per adult per day. It is preferred to administer the above dosage once a day, or twice or three or more times a day.

[0067] In the present invention, other materials having the effects of promoting ammonia metabolism, improving stamina, anti-fatigue, improving cognitive function, or preventing or improving Alzheimer's disease can be used as active ingredients in combination with hesperidins and ornithine. Examples of such other materials include arginine, citrulline, catechins, and combinations thereof.

[0068] The present invention also encompasses the following substances, manufacturing methods, uses, methods, etc. as exemplary embodiments, but the present invention is not limited to these embodiments.

[0069] [1] An ammonia metabolism promoter containing hesperidin and ornithine as active ingredients. [2] An endurance enhancer containing hesperidin and ornithine as active ingredients. [3] An anti-fatigue agent containing hesperidin and ornithine as active ingredients. [4] A cognitive function improving agent containing hesperidin and ornithine as active ingredients. [5] The agent according to [4], wherein the improvement in cognitive function is improvement of cognitive decline accompanied by excessive accumulation of ammonia. [6] The agent according to [4] or [5], wherein the improvement in cognitive function is improvement of cognitive decline associated with Alzheimer's disease. [7] An agent for preventing or ameliorating Alzheimer's disease, comprising hesperidins and ornithine as active ingredients. [8] An agent for preventing or ameliorating hyperammonemia, comprising hesperidins and ornithine as active ingredients. [9] An agent for preventing or improving hepatic encephalopathy, comprising hesperidins and ornithine as active ingredients.

[10] An agent for preventing or improving chronic fatigue syndrome, comprising hesperidins and ornithine as active ingredients.

[11] The agent according to any one of [1] to

[10] , wherein the hesperidins are preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and hesperidin sugar adducts, and more preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and glucosylhesperidin.

[12] The agent according to any one of [1] to

[11] , preferably, the agent is a pharmaceutical or quasi-drug and contains the hesperidins in an amount of 0.1 to 20% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 1 to 20% by mass, 1 to 10% by mass, 1 to 5% by mass, 3 to 20% by mass, 3 to 10% by mass, or 3 to 5% by mass.

[13] The agent according to

[12] , which preferably contains the ornithine in an amount of 0.3 to 60% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 3 to 60% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 5 to 60% by mass, 5 to 30% by mass, 5 to 20% by mass, 5 to 15% by mass, 9 to 60% by mass, 9 to 30% by mass, 9 to 20% by mass, or 9 to 15% by mass.

[14] The agent according to any one of [1] to

[11] , preferably in the form of a food product, containing the hesperidins in an amount of 0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, or 1 to 5% by mass.

[15] The agent according to

[14] , which preferably contains the ornithine in an amount of 0.15 to 30% by mass, 0.15 to 20% by mass, 0.15 to 15% by mass, 0.15 to 5% by mass, 0.15 to 3% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 0.3 to 5% by mass, 0.3 to 3% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 3 to 5% by mass, 5 to 30% by mass, 5 to 20% by mass, or 5 to 15% by mass.

[16] The agent according to any one of [1] to

[15] , wherein the mass ratio of the hesperidins to the ornithine is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, still more preferably 1:1-5, and even more preferably 1:2-4.

[17] The agent according to any one of [1] to

[16] , wherein the agent is preferably an oral agent.

[0070]

[18] Use of hesperidins and ornithine for the production of an ammonia metabolism promoter.

[19] Use of hesperidins and ornithine for the production of an endurance enhancing agent.

[20] Use of hesperidins and ornithine for the production of an anti-fatigue agent.

[21] Use of hesperidins and ornithine for the manufacture of an agent for improving cognitive function.

[22] The use according to

[21] , wherein the improvement of cognitive function is improvement of cognitive decline associated with excessive accumulation of ammonia.

[23] The use according to

[21] or

[22] , wherein the improvement in cognitive function is improvement of cognitive decline associated with Alzheimer's disease.

[24] Use of hesperidins and ornithine for the manufacture of an agent for the prevention or amelioration of Alzheimer's disease.

[25] Use of hesperidins and ornithine for the manufacture of an agent for preventing or ameliorating hyperammonemia.

[26] Use of hesperidins and ornithine for the manufacture of an agent for preventing or ameliorating hepatic encephalopathy.

[27] Use of hesperidins and ornithine for the manufacture of an agent for preventing or ameliorating chronic fatigue syndrome.

[28] The use according to any one of

[18] to

[27] , wherein the hesperidins are preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and hesperidin sugar adducts, and more preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and glucosylhesperidin.

[29] The use according to any one of

[18] to

[28] , preferably wherein the agent is a drug or a quasi-drug and contains the hesperidins in an amount of 0.1 to 20% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 1 to 20% by mass, 1 to 10% by mass, 1 to 5% by mass, 3 to 20% by mass, 3 to 10% by mass, or 3 to 5% by mass.

[30] The use according to

[29] , wherein the agent preferably contains the ornithine in an amount of 0.3 to 60% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 3 to 60% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 5 to 60% by mass, 5 to 30% by mass, 5 to 20% by mass, 5 to 15% by mass, 9 to 60% by mass, 9 to 30% by mass, 9 to 20% by mass, or 9 to 15% by mass.

[31] The use according to any one of

[18] to

[28] , preferably wherein the agent is a food and contains the hesperidins in an amount of 0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, or 1 to 5% by mass.

[32] The use according to

[31] , wherein the agent preferably contains the ornithine in an amount of 0.15 to 30% by mass, 0.15 to 20% by mass, 0.15 to 15% by mass, 0.15 to 5% by mass, 0.15 to 3% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 0.3 to 5% by mass, 0.3 to 3% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 3 to 5% by mass, 5 to 30% by mass, 5 to 20% by mass, or 5 to 15% by mass.

[33] The use according to any one of

[18] to

[32] , wherein the mass ratio of the hesperidins to the ornithine in the agent is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, even more preferably 1:1-5, and still more preferably 1:2-4.

[34] The use according to any one of

[18] to

[33] , wherein the agent is preferably an oral agent.

[0071]

[35] Use of hesperidins and ornithine for promoting ammonia metabolism.

[36] Use of hesperidins and ornithine to improve endurance.

[37] Use of hesperidins and ornithine for anti-fatigue.

[38] Use of hesperidins and ornithine for improving cognitive function.

[39] The use according to

[38] , wherein the improvement of cognitive function is improvement of cognitive decline associated with excessive accumulation of ammonia.

[40] The use described in

[38] or

[39] , wherein the improvement in cognitive function is improvement of cognitive decline associated with Alzheimer's disease.

[41] Use of hesperidins and ornithine for the prevention or amelioration of Alzheimer's disease.

[42] Use of hesperidins and ornithine for the prevention or amelioration of hyperammonemia.

[43] Use of hesperidins and ornithine for the prevention or amelioration of hepatic encephalopathy.

[44] Use of hesperidins and ornithine for the prevention or improvement of chronic fatigue syndrome.

[45] The use according to any one of

[35] to

[44] , wherein the hesperidins are preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and hesperidin sugar adducts, and more preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and glucosylhesperidin.

[46] The use according to any one of

[35] to

[45] , wherein the hesperidins and ornithine are preferably used in the form of a pharmaceutical or quasi-drug containing the hesperidins in an amount of 0.1 to 20% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 1 to 20% by mass, 1 to 10% by mass, 1 to 5% by mass, 3 to 20% by mass, 3 to 10% by mass, or 3 to 5% by mass.

[47] The use according to

[46] , wherein the pharmaceutical or quasi-drug preferably contains the ornithine in an amount of 0.3 to 60% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 3 to 60% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 5 to 60% by mass, 5 to 30% by mass, 5 to 20% by mass, 5 to 15% by mass, 9 to 60% by mass, 9 to 30% by mass, 9 to 20% by mass, or 9 to 15% by mass.

[48] ​​The use according to any one of

[35] to

[45] , wherein the hesperidins and ornithine are preferably used in the form of a food containing the hesperidins in an amount of 0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, or 1 to 5% by mass.

[49] The use according to

[48] , wherein the food preferably contains the ornithine in an amount of 0.15 to 30% by mass, 0.15 to 20% by mass, 0.15 to 15% by mass, 0.15 to 5% by mass, 0.15 to 3% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 0.3 to 5% by mass, 0.3 to 3% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 3 to 5% by mass, 5 to 30% by mass, 5 to 20% by mass, or 5 to 15% by mass.

[50] The use according to any one of

[35] to

[49] , wherein the mass ratio of the hesperidins to the ornithine is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, even more preferably 1:1-5, and still more preferably 1:2-4.

[51] The use according to any one of

[35] to

[50] , wherein the hesperidins and ornithine are preferably administered orally.

[0072]

[52] A combination of hesperidins and ornithine for use in promoting ammonia metabolism.

[53] A combination of hesperidins and ornithine for use in improving endurance.

[54] A combination of hesperidins and ornithine for use in anti-fatigue.

[55] A combination of hesperidins and ornithine for use in improving cognitive function.

[56] The combination of hesperidins and ornithine described in

[55] , wherein the improvement of cognitive function is improvement of cognitive decline accompanied by excessive accumulation of ammonia.

[57] The combination of hesperidins and ornithine according to

[55] or

[56] , wherein the improvement of cognitive function is improvement of cognitive decline associated with Alzheimer's disease.

[58] A combination of hesperidins and ornithine for use in preventing or ameliorating Alzheimer's disease.

[59] A combination of hesperidins and ornithine for use in preventing or ameliorating hyperammonemia.

[60] A combination of hesperidins and ornithine for use in the prevention or amelioration of hepatic encephalopathy.

[61] A combination of hesperidins and ornithine for use in the prevention or amelioration of chronic fatigue syndrome.

[62] The combination of a hesperidin compound and ornithine according to any one of

[52] to

[61] , wherein the hesperidin compound is preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and hesperidin sugar adducts, and more preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and glucosylhesperidin.

[63] The combination of hesperidins and ornithine according to any one of

[52] to

[62] , which is preferably a pharmaceutical or quasi-drug containing the hesperidins in an amount of 0.1 to 20% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 1 to 20% by mass, 1 to 10% by mass, 1 to 5% by mass, 3 to 20% by mass, 3 to 10% by mass, or 3 to 5% by mass.

[64] The combination of hesperidins and ornithine according to

[63] , wherein the pharmaceutical or quasi-drug preferably contains the ornithine in an amount of 0.3 to 60% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 3 to 60% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 5 to 60% by mass, 5 to 30% by mass, 5 to 20% by mass, 5 to 15% by mass, 9 to 60% by mass, 9 to 30% by mass, 9 to 20% by mass, or 9 to 15% by mass.

[65] The combination of hesperidins and ornithine according to any one of

[52] to

[62] , which is preferably a food product containing the hesperidins in an amount of 0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, or 1 to 5% by mass.

[66] The combination of hesperidins and ornithine according to

[65] , wherein the food preferably contains the ornithine in an amount of 0.15 to 30% by mass, 0.15 to 20% by mass, 0.15 to 15% by mass, 0.15 to 5% by mass, 0.15 to 3% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 0.3 to 5% by mass, 0.3 to 3% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 3 to 5% by mass, 5 to 30% by mass, 5 to 20% by mass, or 5 to 15% by mass.

[67] The combination of hesperidins and ornithine according to any one of

[52] to

[66] , wherein the mass ratio of the hesperidins to the ornithine is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, still more preferably 1:1-5, and even more preferably 1:2-4.

[68] The combination of hesperidins and ornithine according to any one of

[52] to

[67] , which is preferably administered orally.

[69] The combination of hesperidins and ornithine according to any one of

[52] to

[68] , wherein the daily dosage of the hesperidins and ornithine for an adult is as follows: [Hesperidins (converted into hesperidin equivalent)] preferably 100 to 3000 mg / 60 kg body weight, more preferably 250 to 2000 mg / 60 kg body weight, even more preferably 250 to 1000 mg / 60 kg body weight, still more preferably 250 to 600 mg / 60 kg body weight; [Ornithine (free form equivalent)] preferably 100 to 5000 mg / 60 kg body weight, more preferably 250 to 3000 mg / 60 kg body weight, even more preferably 400 to 2000 mg / 60 kg body weight, even more preferably 500 to 2000 mg / 60 kg body weight, even more preferably 800 to 1600 mg / 60 kg body weight, or preferably 250 to 800 mg / 60 kg body weight, even more preferably 500 to 800 mg / 60 kg body weight.

[70] The combination of hesperidins and ornithine according to

[69] , wherein the dosages of hesperidins and ornithine are administered once a day, or in divided doses two or three or more times a day.

[0073]

[71] A method for promoting ammonia metabolism, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[72] A method for improving endurance, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[73] An anti-fatigue method, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[74] A method for improving cognitive function, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[75] The method described in

[74] , wherein the improvement of cognitive function is improvement of cognitive decline associated with excessive accumulation of ammonia.

[76] The method described in

[74] or

[75] , wherein the improvement of cognitive function is an improvement of cognitive decline associated with Alzheimer's disease.

[77] A method for preventing or ameliorating Alzheimer's disease, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[78] A method for preventing or ameliorating hyperammonemia, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[79] A method for preventing or ameliorating hepatic encephalopathy, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[80] A method for preventing or ameliorating chronic fatigue syndrome, comprising administering effective amounts of hesperidins and ornithine to a subject in need thereof.

[81] The method according to any one of

[71] to

[80] , wherein the hesperidins are preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and hesperidin sugar adducts, and more preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and glucosylhesperidin.

[82] The method described in any one of

[71] to

[81] , wherein the mass ratio of the hesperidins to the ornithine is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, even more preferably 1:1-5, and still more preferably 1:2-4.

[83] The method according to any one of

[71] to

[82] , wherein the administration is preferably oral administration.

[84] The method according to any one of

[71] to

[83] , wherein the daily dose of the hesperidins and ornithine per adult is as follows: [Hesperidins (converted into hesperidin equivalent)] preferably 100 to 3000 mg / 60 kg body weight, more preferably 250 to 2000 mg / 60 kg body weight, even more preferably 250 to 1000 mg / 60 kg body weight, still more preferably 250 to 600 mg / 60 kg body weight; [Ornithine (free form equivalent)] preferably 100 to 5000 mg / 60 kg body weight, more preferably 250 to 3000 mg / 60 kg body weight, even more preferably 400 to 2000 mg / 60 kg body weight, even more preferably 500 to 2000 mg / 60 kg body weight, even more preferably 800 to 1600 mg / 60 kg body weight, or preferably 250 to 800 mg / 60 kg body weight, even more preferably 500 to 800 mg / 60 kg body weight.

[85] The method described in

[84] , wherein the dosage of hesperidins and ornithine is administered once a day, or in divided doses two or three or more times a day.

[0074]

[86] A food for promoting ammonia metabolism, containing hesperidin and ornithine as active ingredients.

[87] A food for improving endurance containing hesperidin and ornithine as active ingredients.

[88] An anti-fatigue food containing hesperidin and ornithine as active ingredients.

[89] A food for improving cognitive function containing hesperidin and ornithine as active ingredients.

[90] The food described in

[89] , wherein the improvement of cognitive function is the improvement of cognitive decline associated with excessive accumulation of ammonia.

[91] The food described in

[89] or

[90] , wherein the improvement of cognitive function is improvement of cognitive decline associated with Alzheimer's disease.

[92] A food for preventing or improving Alzheimer's disease, comprising hesperidin and ornithine as active ingredients.

[93] A food for preventing or improving hyperammonemia, comprising hesperidins and ornithine as active ingredients.

[94] A food for preventing or improving hepatic encephalopathy, containing hesperidin and ornithine as active ingredients.

[95] A food for preventing or improving chronic fatigue syndrome, containing hesperidin and ornithine as active ingredients.

[96] The food according to any one of

[86] to

[95] , wherein the hesperidins are preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and hesperidin sugar adducts, and more preferably at least one selected from the group consisting of hesperidin, methylhesperidin, and glucosylhesperidin.

[97] The food according to any one of

[86] to

[96] , preferably containing the hesperidins in an amount of 0.05 to 10% by mass, 0.05 to 5% by mass, 0.05 to 1% by mass, 0.1 to 10% by mass, 0.1 to 5% by mass, 0.1 to 1% by mass, 1 to 10% by mass, or 1 to 5% by mass.

[98] The food according to

[97] , preferably containing the ornithine in an amount of 0.15 to 30% by mass, 0.15 to 20% by mass, 0.15 to 15% by mass, 0.15 to 5% by mass, 0.15 to 3% by mass, 0.3 to 30% by mass, 0.3 to 20% by mass, 0.3 to 15% by mass, 0.3 to 5% by mass, 0.3 to 3% by mass, 3 to 30% by mass, 3 to 20% by mass, 3 to 15% by mass, 3 to 5% by mass, 5 to 30% by mass, 5 to 20% by mass, or 5 to 15% by mass.

[99] The food according to any one of

[86] to

[98] , wherein the mass ratio of the hesperidins to the ornithine is preferably 1:0.1-10, more preferably 1:0.5-10, even more preferably 1:1-8, even more preferably 1:1-5, and still more preferably 1:2-4. EXAMPLES

[0075] The present invention will be described in more detail below using examples, although the technical scope of the present invention is not limited to these examples.

[0076] Example 1: Effects of hesperidin and ornithine on ammonia metabolism (urea production) in human hepatocytes Human iPS cell-derived hepatocytes (iCell (registered trademark) hepatocytes 2.0 01279 strain (FUJIFILM Wako Pure Chemical Corporation)) were used as hepatocytes, which were seeded and cultured on a collagen-coated 96-well plate specified by the manufacturer according to the manufacturer's recommended method, and each material was evaluated on the 8th day after seeding. The hepatocytes were cultured for 24 hours at 37°C in a control medium, a hesperidin-containing medium (control medium containing 4.2 μM hesperidin), an ornithine-containing medium (control medium containing 1 mM ornithine hydrochloride), or a hesperidin and ornithine-containing medium (control medium containing 4.2 μM hesperidin and 1 mM ornithine hydrochloride), and then evaluated. Hesperidin (Hamari Chemical Industry Co., Ltd.) was used as hesperidin, and L(+)-ornithine monohydrochloride (FUJIFILM Wako Pure Chemical Corporation) was used as ornithine. The control medium was 5 mM ammonium chloride-containing Williams' Medium E (containing sodium bicarbonate, no phenol red, no glutamine, no FBS, with Glutamax, penicillin / streptomycin, and 100 μM ascorbic acid). After incubation, the culture supernatant was collected and the urea contained therein was quantified using QuantiChrom Urea Assay Kit (100 tests) (BioAssay Systems). A significant difference test was performed between the quantitative values ​​and the control group (Dunnett test, vs. control group, n = 4).

[0077] The quantitative results of urea are shown in Figure 1. Compared to the control group, the urea concentration in the medium in the hesperidin and ornithine-containing groups was statistically significantly increased (*p<0.05), indicating that ammonia metabolism was promoted.

[0078] Example 2: Effects of hesperidins and ornithine on ammonia metabolism (urea production) in human hepatocytes Human iPS cell-derived hepatocytes (iCell (registered trademark) hepatocytes 2.0 01279 strain (FUJIFILM Wako Pure Chemical Industries, Ltd.)) were used as hepatocytes, which were seeded and cultured on a collagen-coated 96-well plate specified by the manufacturer according to the manufacturer's recommended method, and each material was evaluated on the 8th day after seeding. The hepatocytes were cultured for 24 hours at 37°C in a control medium, a medium containing hesperidin and ornithine (a control medium containing 42 μM hesperidin and 1 mM ornithine hydrochloride), and a medium containing methyl hesperidin and ornithine (a control medium containing 42 μM methyl hesperidin and 1 mM ornithine hydrochloride), and then evaluated. Hesperidin (Hamari Chemical Industries, Ltd.) was used as the hesperidin, L(+)-ornithine monohydrochloride (FUJIFILM Wako Pure Chemical Industries, Ltd.) was used as the ornithine, and methyl hesperidin (Hamari Chemical Industries, Ltd.) was used as the methyl hesperidin. The control medium was 5 mM ammonium chloride-containing Williams' Medium E (containing sodium bicarbonate, no phenol red, no glutamine, no FBS, with Glutamax, penicillin / streptomycin, and 100 μM ascorbic acid). After incubation, the culture supernatant was collected and the urea contained therein was quantified using QuantiChrom Urea Assay Kit (100 tests) (BioAssay Systems). A significant difference test was performed between the quantitative values ​​and the control group (Dunnett test, vs. control group, n = 4).

[0079] The quantitative results of urea are shown in Figure 2. Compared to the control group, the urea concentration in the medium was statistically significantly increased (*p<0.05) in the hesperidin and ornithine-containing group and the methylhesperidin and ornithine-containing group, indicating that ammonia metabolism was promoted.

[0080] Reference Example 1: Effect of urea cycle activators on ammonia metabolism (urea production) in human iPS cell-derived hepatocytes Human iPS cell-derived hepatocytes (iCell (registered trademark) hepatocytes 2.0 01279 strain (FUJIFILM Wako Pure Chemical Corporation)) were used as hepatocytes, which were seeded and cultured on a collagen-coated 96-well plate specified by the manufacturer according to the manufacturer's recommended method, and urea cycle activators were evaluated on the 8th day after seeding. The hepatocytes were cultured for 24 hours at 37°C in a control medium or a medium containing a urea cycle activator (control medium containing 1 μM dexamethasone and 50 μM CPT-cAMP (8-(4-chlorophenylthio)adenosine 3'5'-(cyclic)-sodium phosphate)) for evaluation. The control medium used was Williams' Medium E medium containing 5 mM ammonium chloride (containing sodium bicarbonate, no phenol red, no glutamine, no FBS, with Glutamax added, penicillin / streptomycin added, and 100 μM ascorbic acid added). After the culture, the culture supernatant was collected and the urea contained therein was quantified using the QuantiChrom Urea Assay Kit (100 tests) (BioAssay Systems). A significant difference test was performed between the quantitative values ​​and the control group (t-test, n = 4).

[0081] The quantitative results of urea are shown in Figure 3. Compared to the control group, the urea cycle activator-containing group showed a statistically significant (*p<0.05) increase in urea concentration in the medium, indicating that ammonia metabolism was promoted, and it was confirmed that ammonia metabolism can be evaluated using this cell evaluation system.

Claims

1. An ammonia metabolism promoter comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding the case of a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

2. An endurance enhancer comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding the case of a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

3. An anti-fatigue agent comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding the case of a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

4. A cognitive function improving agent comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding the case of a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

5. The agent according to claim 4, wherein the improvement in cognitive function is improvement of cognitive decline accompanied by excessive accumulation of ammonia.

6. The agent according to claim 4 or 5, wherein the improvement in cognitive function is improvement of cognitive decline associated with Alzheimer's disease.

7. An agent for preventing or ameliorating Alzheimer's disease, comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding the case of a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

8. The agent according to any one of claims 1 to 7, wherein the mass ratio of at least one selected from the group consisting of hesperidin and methylhesperidin to ornithine is 1:0.1-10.

9. A food for promoting ammonia metabolism, comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

10. A food for improving endurance, comprising as active ingredients at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine (excluding a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

11. An anti-fatigue food containing at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding the case of a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

12. A food for improving cognitive function, comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

13. The food according to claim 12, wherein the improvement in cognitive function is an improvement in cognitive decline associated with excessive accumulation of ammonia.

14. The food according to claim 12 or 13, wherein the improvement in cognitive function is improvement of cognitive decline associated with Alzheimer's disease.

15. A food for preventing or improving Alzheimer's disease, comprising at least one selected from the group consisting of hesperidin and methylhesperidin, and ornithine as active ingredients (excluding a packaged hesperidin- and ornithine-containing beverage that satisfies any one or more of the following conditions (A) to (D): (A) the concentration of ethyl acetate in the beverage is 0.25 ppm or more; (B) the concentration of ethyl butyrate in the beverage is 0.05 ppm or more; (C) the concentration of methyl caproate in the beverage is 0.55 ppm or more; (D) The concentration of butyl butyrate in the beverage is 0.25 ppm or more.

16. The food according to any one of claims 9 to 15, wherein the mass ratio of at least one selected from the group consisting of hesperidin and methylhesperidin to ornithine is 1:0.1-10.

Images