Dopamine release amount enhancing agent, exercise control function improving agent, exercise learning function improving agent, and activity desire improving agent

Abstract

The purpose of the present invention is to provide a novel dopamine release amount-enhancing agent, exercise control function-improving agent, exercise learning function-improving agent, and activity desire-improving agent. Provided are a dopamine release amount-enhancing agent, a motion control function-improving agent, a motion learning function-improving agent, and an activity desire-improving agent, each of which contains tryptophan-arginine as an active ingredient.

Description

Technical Field

[0001] This invention relates to techniques for enhancing dopamine release, improving motor control, improving motor learning, or improving activity motivation (e.g., dopamine release enhancers, motor control improvers, motor learning improvers, and activity motivation improvers). Background Technology

[0002] It is known that dopamine signaling affects motor control, motor learning, and desire to move (Non-Patent Literature 1 and 2).

[0003] Patent Document 1 discloses a composition comprising the decapeptide LSSTQAQQSY, LSSTQAQQSW, or LSSTQAQQSF for improving low libido, depression, or depressive mood disorders. Patent Document 1 also discloses the manufacture of the decapeptide LSSTQAQQSY by hydrolyzing β-conglycin, a type of soy protein, with a thermophilic protease. It should be noted that "L" represents lysine, "S" represents serine, "T" represents threonine, "Q" represents glutamine, "A" represents alanine, "Y" represents tyrosine, "W" represents tryptophan, and "F" represents phenylalanine.

[0004] Patent Document 2 discloses a composition for improving, maintaining, and / or enhancing the motor control and / or motor learning functions of the tetrapeptide GTWY. Patent Document 2 also discloses the production of the peptide GTWY via enzymatic hydrolysis of whey protein. It should be noted that "G" represents glycine, "T" represents threonine, "W" represents tryptophan, and "Y" represents tyrosine.

[0005] Patent Document 3 discloses a composition for alleviating or preventing lack of desire / motivation and / or low vitality, comprising dipeptides LH, DV, or MH. Patent Document 3 describes that the effects of these dipeptides are exerted by inhibiting microglial inflammation. Patent Document 3 also describes the production of a peptide extract containing dipeptides LH, DV, or MH by dissolving skim milk powder or skim soy protein in water and subjecting it to enzymatic treatment. It should be noted that "L" represents lysine, "H" represents histidine, "D" represents aspartic acid, "V" represents valine, and "M" represents methionine.

[0006] Existing technical documents

[0007] Patent documents

[0008] Patent Document 1: Japanese Patent Application Publication No. 2020-040982

[0009] Patent Document 2: Japanese Patent Application Publication No. 2020-002078

[0010] Patent Document 3: Japanese Patent Application Publication No. 2016-222646

[0011] Non-patent literature

[0012] Non-patent literature 1: Clark, BC, et al., The Aging Brain&the Dorsal BasalGanglia: Implications for Age-Related Limitations of Mobility., Adv.Geriatr.Med.Res., 2019.1:e190008.

[0013] Non-patent literature 2: Schultz, W., Predictive reward signal of dopamine neurons. J. Neurophysiol., 1998. 80(1): p. 1-27. Summary of the Invention

[0014] The problem the invention aims to solve

[0015] The purpose of this invention is to provide new technologies (e.g., novel dopamine release enhancers, motor control function improvers, motor learning function improvers, and activity desire improvers) for enhancing dopamine release, improving motor control function, improving motor learning function, or improving activity desire.

[0016] Solution for solving the problem

[0017] The present invention provides the following formulation.

[0018] [A1] A dopamine release enhancer containing tryptophan arginine as the active ingredient.

[0019] [A2] A motor control function improver containing tryptophan arginine as an active ingredient.

[0020] [A3] According to the motor control function improver described in [A2], the motor control function improver is used to improve one or more motor control functions selected from motor coordination function, balance function, reaction ability and fine motor skills.

[0021] [A4] A motor learning function improver containing tryptophan arginine as an active ingredient.

[0022] [A5] An activity motivation enhancer containing tryptophan arginine as its active ingredient.

[0023] In addition, the present invention provides the following uses.

[0024] [B1] Use of tryptophan arginine in the manufacture of dopamine release enhancers.

[0025] [B2] Use of tryptophanyl arginine in the manufacture of a motor control improver.

[0026] [B3] According to the use described in [B2], the motor control function improver is used to improve one or more motor control functions selected from motor coordination, balance, reaction ability and fine motor skills.

[0027] [B4] Use of tryptophanyl arginine in the manufacture of a motor learning function improver.

[0028] [B5] Use of tryptophanyl arginine in the manufacture of a desire-enhancing agent.

[0029] In addition, the present invention provides the following method.

[0030] [C1] A method for enhancing dopamine release, comprising the step of administering tryptophan arginine to a subject requiring enhanced dopamine release.

[0031] [C2] A method for improving motor control function, comprising the step of administering tryptophan arginine to an object for which motor control function needs to be improved.

[0032] [C3] According to the method described in [C2], improving motor control function means improving one or more motor control functions selected from motor coordination function, balance function, reaction ability and fine motor skills.

[0033] [C4] A method for improving motor learning function, comprising the step of administering tryptophan-arginine to an individual for whom motor learning function needs to be improved.

[0034] [C5] A method for improving activity desire, comprising the step of administering tryptophan-arginine to an object for which activity desire needs to be improved.

[0035] In addition, the present invention provides the following uses.

[0036] [D1] The use of tryptophan arginine as an active ingredient in dopamine release enhancers.

[0037] [D2] The use of tryptophan arginine as an effective ingredient in a motor control improver.

[0038] [D3] According to the use described in [D2], the motor control function improver is used to improve one or more motor control functions selected from motor coordination, balance, reaction ability and fine motor skills.

[0039] [D4] The use of tryptophan arginine as an effective ingredient in an exercise learning function improver.

[0040] [D5] Use of tryptophan arginine as an active ingredient in an activity motivation enhancer.

[0041] The effects of the invention

[0042] According to the present invention, new technologies (e.g., novel dopamine release enhancers, motor control function improvers, motor learning function improvers, or activity desire improvers) are provided for enhancing dopamine release, improving motor control function, improving motor learning function, or improving activity desire. Detailed Implementation

[0043] The present invention will now be described.

[0044] The present invention provides a dopamine release enhancer (hereinafter sometimes referred to as the "first formulation"), a motor control function improver (hereinafter sometimes referred to as the "second formulation"), a motor learning function improver (hereinafter sometimes referred to as the "third formulation"), and an activity desire improver (hereinafter sometimes referred to as the "fourth formulation").

[0045] Active Ingredients

[0046] The first to fourth formulations contain tryptophan arginine as an active ingredient. Tryptophan arginine is a dipeptide composed of the amino acid sequence shown in WR. "W" represents tryptophan, and "R" represents arginine. Hereinafter, tryptophan arginine will sometimes be referred to as "the dipeptide of the present invention".

[0047] The dipeptides of the present invention can be obtained through chemical synthesis or through chemical decomposition or enzymatic hydrolysis of proteins. Examples of proteins include those derived from soybeans (e.g., soybean glycinin). The safety of the dipeptides of the present invention is supported by extensive human experience with soybeans as a representative example.

[0048] The first to fourth formulations may or may not contain peptides other than the dipeptides of the present invention, obtained by chemically decomposing or enzymatically hydrolyzing proteins (e.g., proteins derived from soybeans). In one embodiment, the first to fourth formulations do not contain peptides other than the dipeptides of the present invention, obtained by chemically decomposing or enzymatically hydrolyzing proteins (e.g., proteins derived from soybeans). Examples of peptides other than the dipeptides of the present invention obtained by chemically decomposing or enzymatically hydrolyzing proteins (e.g., proteins derived from soybeans) include, for example, dipeptides WM, LH, DV or MH (Japanese Patent Application Laid-Open No. 2016-222646), decapeptides LSSTQAQQSY (Japanese Patent Application Laid-Open No. 2020-40982), YPFVV (Japanese Patent Application Laid-Open No. 2007-91656), etc. It should be noted that "W" represents tryptophan, "M" represents methionine, "L" represents lysine, "H" represents histidine, "D" represents aspartic acid, "V" represents valine, "S" represents serine, "T" represents threonine, "Q" represents glutamine, "A" represents alanine, "Y" represents tyrosine, "P" represents proline, and "F" represents phenylalanine.

[0049] The first to fourth formulations may or may not contain any active ingredient other than the dipeptide of the present invention. In one embodiment, the first to fourth formulations contain the dipeptide of the present invention as the sole active ingredient (i.e., the first to fourth formulations do not contain any active ingredient other than the dipeptide of the present invention). Examples of peptides not included in the first to fourth formulations include, for example, dipeptides LH, DV, or MH (Japanese Patent Application Laid-Open No. 2016-222646), tetrapeptide GTWY (Japanese Patent Application Laid-Open No. 2020-002078), and decapeptides LSSTQAQQSY, LSSTQAQQSW, or LSSTQAQQSF (Japanese Patent Application Laid-Open No. 2020-40982). It should be noted that "L" represents lysine, "H" represents histidine, "D" represents aspartic acid, "V" represents valine, "M" represents methionine, "G" represents glycine, "T" represents threonine, "W" represents tryptophan, "Y" represents tyrosine, "S" represents serine, "Q" represents glutamine, "A" represents alanine, and "F" represents phenylalanine.

[0050] The dipeptide of the present invention may be composed of only L-type amino acids, only D-type amino acids, or both L-type and D-type amino acids.

[0051] The dipeptide of the present invention can be in the form of a pharmaceutically acceptable salt or solvate. The phrase "the first to fourth formulations contain tryptophanyl arginine as an active ingredient" includes not only the case where the first to fourth formulations contain tryptophanyl arginine itself, but also the case where the first to fourth formulations contain tryptophanyl arginine in the form of a pharmaceutically acceptable salt and / or solvate. Examples of pharmaceutically acceptable salts include acid addition salts, metal salts, ammonium salts, and organic amine addition salts. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, nitrate, sulfate, and phosphate; and organic acid salts such as acetate, oxalate, maleate, fumarate, citrate, benzoate, and methanesulfonate. Examples of pharmaceutically acceptable metal salts include alkali metal salts such as sodium and potassium salts; alkaline earth metal salts such as magnesium and calcium salts; and aluminum and zinc salts. Examples of pharmaceutically acceptable ammonium salts include ammonium and tetramethylammonium salts. Examples of pharmaceutically acceptable addition salts of organic amines include morpholine and piperidine.

[0052] The content of the dipeptide of the present invention in the first to fourth formulations can be appropriately adjusted considering the form of each formulation, the frequency of daily administration of each formulation, and the daily dosage of each formulation. The first to fourth formulations may consist solely of the dipeptide of the present invention. The content of the dipeptide of the present invention in the first to fourth formulations is based on the mass of each of the first to fourth formulations, and for example, can be adjusted to 0.0001% by mass or more and 100% by mass or less, preferably 0.001% by mass or more and 100% by mass or less. The content of the dipeptide of the present invention can be determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS). LC-MS / MS can be performed according to the conditions described in the examples.

[0053] The recipient

[0054] The preferred subjects for administering the dipeptide or the first to fourth formulations of the present invention are mammals such as humans, mice, rats, hamsters, rabbits, dogs, cats, pigs, cattle, horses, and monkeys, with humans being more preferred.

[0055] "use"

[0056] <Primary Use>

[0057] The dipeptide of the present invention has a dopamine release-enhancing effect. That is, the dipeptide of the present invention can enhance the dopamine release. Therefore, the dipeptide of the present invention is useful as an effective ingredient of a dopamine release enhancer. In addition, the dipeptide of the present invention can be given to objects that require enhanced dopamine release in order to enhance dopamine release.

[0058] The first formulation can enhance dopamine release through the dopamine release-enhancing effect of the dipeptide of the present invention.

[0059] "Enhancing dopamine release" refers to increasing the amount of dopamine released (secreted) from dopaminergic neurons in the brain. Examples of dopaminergic neurons include the A8, A9, A10, A11, A12, A13, A14, A15, A16, and A17 cell groups. Cell groups A8-A10 are located in the midbrain, A11-A15 in the diencephalon, A16 in the olfactory bulb, and A17 in the retina.

[0060] Enhancing dopamine release can be achieved by promoting the release of dopamine from dopaminergic neurons in the brain and / or promoting the production of dopamine from dopaminergic neurons in the brain. Therefore, the dipeptide of the present invention is useful as an effective component of a dopamine release promoter and / or a dopamine production promoter. Furthermore, the dipeptide of the present invention can be administered to objects requiring promotion of dopamine release and / or dopamine production.

[0061] By enhancing dopamine release, the amount of dopamine in the brain can be increased. Therefore, the dipeptide of the present invention is useful as an active ingredient in a dopamine-enhancing agent for the brain. Furthermore, the dipeptide of the present invention can be administered to subjects who require increased dopamine levels in the brain.

[0062] The preferred recipients of the dipeptide or first formulation of the present invention are individuals who require enhanced dopamine release.

[0063] In one embodiment, the recipient is a healthy person. In this embodiment, the dipeptide or first formulation of the present invention can prevent a decrease in dopamine release or symptoms or diseases caused by a decrease in dopamine release by enhancing the dopamine release in the recipient. The risk of a decrease in dopamine release may increase with age. Therefore, in a preferred embodiment, the recipient is a healthy person in menopause or old age. Menopause refers to the age at which menopausal symptoms appear, for example, 45 to 55 years of age. Old age refers to advanced age after menopause, for example, 56 years of age or older, preferably 60 years of age or older, and more preferably 65 years of age or older.

[0064] In another embodiment, the recipient is a person experiencing or suspected of experiencing a decrease in dopamine release. In this embodiment, the dipeptide or first formulation of the present invention can prevent or improve a decrease in dopamine release or symptoms or diseases caused by a decrease in dopamine release by enhancing the dopamine release of the recipient. The risk of a decrease in dopamine release may increase with age. Therefore, in a preferred embodiment, the recipient is a person in menopause or old age who experiences or is suspected of experiencing a decrease in dopamine release. The meanings of "menopause" and "old age" are the same as described above.

[0065] "Enhancing dopamine release" includes increasing dopamine levels compared to the subject's current state and increasing dopamine levels compared to healthy individuals of the same age. "Decreased dopamine release" includes decreasing dopamine levels compared to the subject's current state and decreasing dopamine levels compared to healthy individuals of the same age. "Preventing decreased dopamine release" includes maintaining dopamine levels, inhibiting decreased dopamine release, and delaying decreased dopamine release. "Improving decreased dopamine release" includes restoring decreased dopamine release to normal levels. "Preventing symptoms or diseases" includes suppressing the occurrence of symptoms or diseases and delaying the occurrence of symptoms or diseases. "Improving symptoms or diseases" includes suppressing the development or worsening of symptoms or diseases, delaying the development or worsening of symptoms or diseases, alleviating symptoms or diseases, and curing symptoms or diseases. Examples of "symptoms or diseases caused by decreased dopamine release" include limb tremors, postural instability, cognitive impairment, and depressive symptoms.

[0066] Secondary Use

[0067] The dipeptide of the present invention has an effect on improving motor control function. That is, the dipeptide of the present invention can improve motor control function. Therefore, the dipeptide of the present invention is useful as an effective ingredient in a motor control function improver. In addition, the dipeptide of the present invention can be given to subjects who need to improve motor control function. It should be noted that the effect of the dipeptide of the present invention on improving motor control function can be exerted through the dopamine release enhancement effect of the dipeptide of the present invention, but the mechanism of action for improving motor control function is not limited to this.

[0068] The second formulation can improve motor control function through the motor control function-improving effect of the dipeptide of the present invention.

[0069] "Movement" refers to actions produced by voluntary movement (e.g., actions based on voluntary muscles such as limbs and eyes), including daily actions, sports, playing musical instruments, manual operations (without restriction on whether props are used), etc.

[0070] "Motor control function" refers to the brain's ability to consciously and precisely control the movement of skeletal muscles (voluntary movement) to achieve the target action. In other words, it is the brain function that enables skeletal muscle movement to achieve the target action. It mainly includes feedforward control that generates motor commands based on the target state, and feedback control that generates motor commands by using the error signal between the target state and the perceived state.

[0071] Examples of motor control functions include motor coordination, balance, reaction time, and fine motor skills. The second formulation is preferably used to improve one or more motor control functions selected from motor coordination, balance, reaction time, and fine motor skills.

[0072] "Motor coordination function" refers to the ability to coordinate the timing and intensity of multiple muscle groups to perform precise and smooth movements.

[0073] "Balance function" refers to the ability to maintain proper body balance based on information such as vision and inner ear sensation.

[0074] "Responsiveness" refers to the ability to respond quickly to applied stimuli.

[0075] "Fineness" refers to the ability to perform fine movements using the body, especially the hands and fingertips.

[0076] The preferred recipients of the dipeptide or second formulation of the present invention are individuals who require improved motor control.

[0077] In one embodiment, the recipient is a healthy individual. In this embodiment, the dipeptide or second formulation of the present invention can prevent a decline in the recipient's motor control function by maintaining or improving it. "Improving motor control function" in this invention includes maintaining motor control function, enhancing motor control function, and preventing a decline in motor control function. The risk of a decline in motor control function may increase with age. Therefore, in a preferred embodiment, the recipient is a healthy individual in menopause or old age. The meanings of "menopause" and "old age" are the same as described above.

[0078] In another embodiment, the recipient is a person experiencing or suspected of experiencing a decline in motor control function. In this embodiment, the dipeptide or second formulation of the present invention can prevent or improve the decline in motor control function by maintaining or enhancing the motor control function of the recipient. "Improving motor control function" in the present invention includes maintaining motor control function, enhancing motor control function, preventing a decline in motor control function, and improving a decline in motor control function. The risk of a decline in motor control function may increase with age. Therefore, in a preferred embodiment, the recipient is a person in menopause or old age who experiences or is suspected of experiencing a decline in motor control function. The meanings of "menopause" and "old age" are the same as described above.

[0079] "Improving motor control function" includes improving compared to the subject's current state and improving compared to healthy individuals of the same age. "Deteriorating motor control function" includes declining compared to the subject's current state and declining compared to healthy individuals of the same age. "Preventing declining motor control function" includes maintaining motor control function, inhibiting declining motor control function, and delaying declining motor control function. "Improving declining motor control function" includes restoring already diminished motor control function and restoring already diminished motor control function to normal.

[0080] <Third Use>

[0081] The dipeptide of the present invention has an effect on improving motor learning function. That is, the dipeptide of the present invention can improve motor learning function. Therefore, the dipeptide of the present invention is useful as an effective ingredient in a motor learning function improver. In addition, the dipeptide of the present invention can be given to subjects who need to improve their motor learning function. It should be noted that the effect of the dipeptide of the present invention on improving motor learning function can be exerted through the dopamine release enhancement effect of the dipeptide of the present invention, but the mechanism of action for improving motor learning function is not limited to this.

[0082] The third formulation can improve motor learning function through the motor learning function-improving effect of the dipeptide of the present invention.

[0083] "Learning function" refers to the function of changing behavior through environmental stimuli, that is, the function of changing behavior through experience, practice, etc. "Motor learning function" is the motor aspect of learning function.

[0084] As a motor learning function, motor memory can be cited as an example. "Motor memory" is a type of latent memory (unconscious memory that automatically appears with almost no conscious processing). It refers to a function characterized by repeated, progressive learning; that is, the function of storing motor control information acquired through experience and practice as an internal model of movement. In other words, motor memory stores information as motor control information in the brain, and motor learning accumulates this information, thereby achieving continuous updating (refinement, improvement) of motor skills.

[0085] The preferred recipients of the dipeptide or third formulation of the present invention are individuals who require improvement in motor learning function.

[0086] In one embodiment, the recipient is a healthy individual. In this embodiment, the dipeptide or third formulation of the present invention can prevent a decline in the recipient's motor learning function by maintaining or improving it. "Improving motor learning function" in this invention includes maintaining motor learning function, enhancing motor learning function, and preventing a decline in motor learning function. The risk of a decline in motor learning function may increase with age. Therefore, in a preferred embodiment, the recipient is a healthy individual in menopause or old age. The meanings of "menopause" and "old age" are the same as described above.

[0087] In another embodiment, the recipient is a person experiencing or suspected of experiencing a decline in motor learning function. In this embodiment, the dipeptide or third formulation of the present invention can prevent or improve the decline in motor learning function by maintaining or enhancing the motor learning function of the recipient. "Improving motor learning function" in the present invention includes maintaining motor learning function, enhancing motor learning function, preventing a decline in motor learning function, and improving a decline in motor learning function. The risk of a decline in motor learning function may increase with age. Therefore, in a preferred embodiment, the recipient is a person in menopause or old age who experiences or is suspected of experiencing a decline in motor learning function. The meanings of "menopause" and "old age" are the same as described above.

[0088] "Improving motor learning function" includes improving compared to the subject's current state and improving compared to healthy individuals of the same age. "Deteriorating motor learning function" includes declining compared to the subject's current state and declining compared to healthy individuals of the same age. "Preventing declining motor learning function" includes maintaining motor learning function, inhibiting declining motor learning function, and delaying declining motor learning function. "Improving declining motor learning function" includes restoring already diminished motor learning function and restoring already diminished motor learning function to normal.

[0089] <Fourth Use>

[0090] The dipeptide of the present invention has an activity desire-enhancing effect. That is, the dipeptide of the present invention can improve activity desire. Therefore, the dipeptide of the present invention is useful as an active ingredient of an activity desire-enhancing agent. In addition, the dipeptide of the present invention can be given to subjects who need to improve their activity desire. It should be noted that the activity desire-enhancing effect of the dipeptide of the present invention can be exerted through the dopamine release-enhancing effect of the dipeptide of the present invention, but the mechanism of action for improving activity desire is not limited to this.

[0091] The fourth formulation can improve activity desire through the activity desire-enhancing effect of the dipeptide of the present invention.

[0092] "Activity" refers to the movement of the motor organs that consumes more energy than the resting state, including daily activities (such as housework, labor, commuting to school, etc.) and exercise (such as physical activities that are intentionally or planned to maintain or improve physical fitness). "Desire to be active" refers to the mental and psychological state of actively engaging in the above-mentioned "activities".

[0093] Increased activity levels are characterized by higher frequency and duration of exercise, as well as greater energy expenditure during activity. Therefore, increased activity levels can be confirmed by an increase in activity levels from waking to bedtime. Activity levels can be measured as the total time or total energy spent on spontaneous activity from waking to bedtime.

[0094] The preferred recipients of the dipeptide or fourth formulation of the present invention are individuals who require improved activity levels.

[0095] In one embodiment, the recipient is a healthy person. In this embodiment, the dipeptide or fourth formulation of the present invention can prevent a decrease in the recipient's desire to move by maintaining or increasing that desire. "Improving desire to move" in the present invention includes maintaining desire to move, increasing desire to move, and preventing a decrease in desire to move. The risk of a decrease in desire to move may increase with age. Therefore, in a preferred embodiment, the recipient is a healthy person in menopause or old age. The meanings of "menopause" and "old age" are the same as described above.

[0096] In another embodiment, the recipient is a person experiencing or suspected of experiencing a decreased desire to move. In this embodiment, the dipeptide or fourth formulation of the present invention can prevent or improve a decreased desire to move by maintaining or increasing the recipient's desire to move. "Improving desire to move" in the present invention includes maintaining desire to move, increasing desire to move, preventing a decrease in desire to move, and improving a decrease in desire to move. The risk of decreased desire to move may increase with age. Therefore, in a preferred embodiment, the recipient is a person in menopause or old age who experiences or is suspected of experiencing a decreased desire to move. The meanings of "menopause" and "old age" are the same as described above.

[0097] "Increasing the desire to engage in activities" includes increasing it compared to the recipient's current state and compared to healthy individuals of the same age. "Decreased desire to engage in activities" includes decreasing it compared to the recipient's current state and compared to healthy individuals of the same age. "Preventing decreased desire to engage in activities" includes maintaining, inhibiting, and delaying the decline in desire to engage in activities. "Improving decreased desire to engage in activities" includes restoring already decreased desire to engage in activities and restoring already decreased desire to normal levels.

[0098] According to the fourth formulation, it can bring about favorable effects on the mental and psychological aspects of activity, and therefore is expected to prevent or improve depressive symptoms or depression that occur after menopause, especially in old age. In addition, the fourth formulation can increase activity levels, and therefore can incidentally inhibit musculoskeletal decline, potentially reducing the risk of needing nursing care.

[0099] The fourth formulation is highly effective during the first half of the day's activity period. Therefore, it can be used to increase activity levels during this period. The "activity period" refers to the time of day when energy expenditure is relatively high, corresponding to the time from waking to bedtime when daily activities are performed—that is, daytime. Thus, the fourth formulation can be used to increase activity levels during the first half of the day. Based on this ability to increase activity levels during the first half of the day, the fourth formulation may also help prevent or improve insomnia that occurs after menopause, particularly in old age.

[0100] <Usage and Dosage>

[0101] The daily dosage of the dipeptide or the first to fourth formulations of the present invention can be appropriately adjusted taking into account the subject's gender, age, weight, symptoms, etc. For example, the daily dosage of the dipeptide of the present invention for adults is 0.01 to 100 mg, preferably 0.1 to 10 mg.

[0102] The administration time, interval, and daily frequency of the dipeptide or the first to fourth formulations of the present invention can be appropriately adjusted considering the subject's gender, age, weight, symptoms, etc. For example, the dipeptide or the first to fourth formulations of the present invention may be administered daily, every other day, or every two days for a period of one week or more; preferably daily, every other day, or every two days for a period of two weeks or more; more preferably daily, every other day, or every two days for a period of four weeks or more. The daily frequency is not particularly limited and is typically once, twice, or three times.

[0103] The administration route of the dipeptide or the first to fourth formulations of the present invention is not particularly limited as long as it can achieve the target effect (enhanced dopamine release, improved motor control function, improved motor learning function, improved desire to exercise, etc.). In one embodiment, the administration route is oral. In another embodiment, the administration route is non-oral. Examples of non-oral administration routes include intranasal, ophthalmic, ocular, transdermal, intra-respiratory, rectal, intraurinary, subcutaneous, intramuscular, and intravenous administration. Oral administration is preferred.

[0104] <Formulation>

[0105] The first to fourth formulations may consist solely of the dipeptides of the present invention, but are preferably in the form of a composition, more preferably in the form of a food or beverage composition or a pharmaceutical composition.

[0106] Examples of food and beverage combinations include health foods, nutritional supplements, functional foods, health-functional foods (such as foods for specific health purposes, nutritional functional foods, and foods with functional labeling), and foods for special purposes (such as milk powder for pregnant and lactating women).

[0107] Examples of the forms in which food and beverage compositions can be made include solid, powder, paste, semi-liquid, gel, and liquid.

[0108] As a food and beverage composition, there are no particular limitations on any food or beverage containing the dipeptide of the present invention. Examples include instant noodles, steamed or boiled foods, canned foods, microwaveable foods, instant soups / miso soups, freeze-dried foods, and other instant food products; beverages such as soft drinks, fruit juices, vegetable drinks, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, and alcoholic beverages; wheat flour products such as bread, pasta, noodles, cake flour, and bread flour; and maltose, caramel, chewing gum, chocolate, cookies, biscuits, biscuit bars, cakes, and pies. Snacks, crackers, Japanese sweets, mousses, desserts, and other confectionery; sauces, tomato-based seasonings, flavorings, cooking mixes, sauces, broths, curry / Japanese stew sauces, and other condiments; processed oils, butter, margarine, mayonnaise, and other oils; dairy products, including milk beverages, fermented milk, cheese, yogurt, lactic acid bacteria beverages, milk beverages, cheese, ice cream, cream, formula milk powder, liquid milk, and solid milk; canned agricultural products, jams / orange peel jams, cereals, and other processed agricultural products; frozen foods, liquid foods, supplements, etc. The food and beverage compositions, except for containing the dipeptide of this invention, can be manufactured according to conventional manufacturing methods.

[0109] The components contained in the food and beverage composition, other than the dipeptide of the present invention, are not particularly limited. Examples of components other than the dipeptide of the present invention include water, proteins, carbohydrates, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, and flavoring agents. Examples of proteins include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, α-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, lactoferrin, soy protein, egg protein, meat protein, and other animal and plant proteins, their hydrolysates, butter, whey minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids, lactose, and various other milk-derived components. Examples of carbohydrates include common sugars, modified starches (dextrin, soluble starch, British starch, oxidized starch, starch esters, starch ethers, etc.), and dietary fiber. Examples of lipids include lard, fish oil, and their extracted oils, hydrogenated oils, and transesterified oils (animal fats); and palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, and their extracted oils, hydrogenated oils, and transesterified oils (vegetable fats). Examples of vitamins include vitamin A, carotene, B vitamins, vitamin C, vitamin D, vitamin E, vitamin K, vitamin P, vitamin Q, niacin, niacin, pantothenic acid, biotin, inositol, choline, and folic acid. Examples of minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium, and whey minerals. Examples of organic acids include malic acid, citric acid, lactic acid, and tartaric acid. These components can be used alone or in combination of two or more.

[0110] The pharmaceutical composition comprises the dipeptide of the present invention and one or more pharmaceutically acceptable carriers. The pharmaceutical composition is preferably an orally administered pharmaceutical composition (oral dosage form). Examples of pharmaceutical compositions include granules, powders, tablets (including sugar-coated tablets), pills, capsules, syrups, emulsions, suspensions, etc. These formulations can be manufactured using conventional methods with pharmaceutically acceptable carriers. Examples of pharmaceutically acceptable carriers include excipients, binders, diluents, additives, flavorings, buffers, thickeners, colorants, stabilizers, emulsifiers, dispersants, suspending agents, preservatives, etc. A pharmaceutically acceptable carrier can be used alone or in combination of two or more.

[0111] The first to fourth preparations can be packaged in a manner that facilitates the administration of a single dose. The single dose can be one package or multiple packages. When provided in a packaged form, in order to facilitate continuous intake, it is preferably provided in a set form for a certain period of intake (e.g., the intake for several days). The packaging form is not particularly limited as long as it defines a certain amount, and examples include wrapping paper, bags, flexible bags, paper containers, cans, bottles, capsules, etc.

[0112] The uses, dosages, administration methods, etc. of the first to fourth preparations can be indicated on the packaging, container, or accompanying documents (instruction manuals, brochures, promotional materials, brand websites, etc.). Here, "indication" includes all indications used to make the needy person aware of the above-mentioned uses. Such indication can be any indication that can make people associate / analogize the above-mentioned uses, dosages, administration methods, etc., regardless of the purpose of the indication, the content of the indication, the object of the indication, the medium, etc., and can include all indications.

[0113] Examples

[0114] Hereinafter, the present invention will be described in detail based on examples, but the present invention is not limited to the examples.

[0115] Examples and Comparative Examples

[0116] Materials

[0117] PC12 cells were obtained from the RIKEN Institute (Saitama, Japan) and used within 20 passages.

[0118] DMEM medium (Dulbecco's Modified Eagle Medium), penicillin, and streptomycin were purchased from gibco (Carlsbad, CA), FBS (fetal bovine serum) was purchased from Biological Industries (Kibbutz Beit-Haemek, Israel), and HS (horse serum) was purchased from Gibco (Carlsbad, CA).

[0119] Dipeptides (tryptophanyl-arginine (WR), tryptophanyl-phenylalanine (WF), tryptophanyl-histidine (WH), tryptophanyl-methionine (WM)) were purchased from genscript (Piscataway, NJ). All dipeptides are synthetic dipeptides. "W" represents tryptophan, "R" represents arginine, "F" represents phenylalanine, "H" represents histidine, and "M" represents methionine.

[0120] Culture of PC12 Cells

[0121] PC12 cells were cultured in 96-well plates at a concentration of 2 × 10⁻⁶ m³ / well using DMEM containing 10% HS (horse serum), 10% FBS (fetal bovine serum), 100 U / mL penicillin, and 100 μg / mL streptomycin. 4 Seed cells per well and cultured at 5% CO2 at 37°C. Culture for 3–5 days, and use for experiments after confirming confluence.

[0122] <Adding dipeptides to PC12 cells>

[0123] The culture medium was removed from PC12 cells, and the cells were washed twice with PBS (phosphate-buffered saline). The wells of a 96-well plate were divided into five groups: no stimulation (5 wells), WF treatment (3 wells), WH treatment (3 wells), WM treatment (3 wells), and WR treatment (3 wells). The WR treatment group corresponds to the example case, while the no stimulation, WF, WH, and WM treatment groups correspond to the comparative cases.

[0124] DMSO (dimethyl sulfoxide) was added to the same medium used for culturing PC12 cells to prepare a medium containing 1% by mass DMSO. Dipeptide WF was added to the same medium used for culturing PC12 cells to prepare a medium containing 1 mM dipeptide WF. Similarly, media containing 1 mM dipeptide WH, 1 mM dipeptide WM, and 1 mM dipeptide WR were prepared. The unstimulated group was given a medium containing 1% by volume DMSO, the WF group was given a medium containing 1 mM dipeptide WF, the WH group was given a medium containing 1 mM dipeptide WH, the WM group was given a medium containing 1 mM dipeptide WM, and the WR group was given a medium containing 1 mM dipeptide WR. The cells were incubated at 37°C for 6 hours. After incubation, the supernatant from each well was collected.

[0125] <Sample Preparation>

[0126] Transfer all supernatants to ice. Add 25 μL of 2M perchloric acid to each supernatant to remove protein. After 10 minutes, add 50 μL of 1M sodium acetate for neutralization. After 15 minutes, centrifuge at 4°C and 20000G for 20 minutes. Collect 100 μL of the supernatant, add 10 μL of 2mM 3,4-dihydroxybenzylamine (DHBA) as an internal standard, and then pass the sample through a chromatographic disc (Chromatodisc). The obtained sample is then used for liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0127] <lc-ms ms>

[0128] LC-MS / MS was performed using a SCIEX Triple TOF 6600+ system under the following conditions. A standard curve was prepared using the pure sample, and the peak area of ​​dopamine was corrected using the peak area of ​​the internal standard (DHBA). The concentration of dopamine in the sample (nM) was then calculated based on the standard curve.

[0129] Instrument Name: Triple TOF 6600+ System (SCIEX, Massachusetts, USA)

[0130] Column: Inertsil (registered trademark) ODS-4 (GL Sciences, Tokyo, Japan)

[0131] Eluent A: 0.1% formic acid aqueous solution

[0132] Eluent B: 0.1% formic acid in methanol solution

[0133] Gradient conditions: as recorded in Table 1.

[0134] [Table 1]

[0135] Table 1

[0136] Time (minutes) Eluent A (volume %) Eluent B (volume %) 0-10 minutes 98％ 2％ 11-12 minutes 2％ 98％ 13-14 minutes 98％ 2％

[0137] m / z values ​​of dopamine: Q1 154.1, Q3 90.9

[0138] m / z values ​​of the internal standard (DHBA): Q1 140.1, Q3 123.0

[0139] MS conditions:

[0140] Declustering potential: 37V

[0141] Collision energy: 8V

[0142] Curtain gas: 20 psi

[0143] Gas 1:50psi

[0144] Gas 2: 50psi

[0145] Ion spray voltage: 5500V

[0146] It should be noted that "Q1" refers to the precursor ion, and "Q3" refers to the product ion generated by the collision of the precursor ion with the gas. That is, for dopamine, a precursor ion with m / z of 154.1 is selected, and after collision with the gas, a product ion with m / z of 90.9 is detected. For the internal standard (DHBA), a precursor ion with m / z of 140.1 is selected, and after collision with the gas, a product ion with m / z of 123.0 is detected.

[0147] <Statistical Processing>

[0148] Data are expressed as mean ± standard deviation. For statistical processing, Student's t-test was used to compare the no-stimulation group with each administered group. The significance level was set at 5%.

[0149] <Results>

[0150] The results of the dopamine concentration measurements in the samples are shown in Table 2. As shown in Table 2, a significant increase in dopamine concentration was observed in the WR-treated group compared with the unstimulated group (p<0.05), but no such significant increase in dopamine concentration was observed in the WF-treated, WH-treated, and WM-treated groups.

[0151] [Table 2]

[0152]

Claims

1. A dopamine release enhancer containing tryptophan arginine as an active ingredient.

2. A motor control function improver containing tryptophan arginine as an active ingredient.

3. The motion control function improver according to claim 2, wherein, The motor control function improver is used to improve one or more motor control functions selected from motor coordination, balance, reaction ability, and fine motor skills.

4. A motor learning function improver containing tryptophan arginine as an active ingredient.

5. An activity motivation enhancer containing tryptophan arginine as an active ingredient.

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