JP7679339B2 - Composition for enhancing the effect of exercise on reducing body fat - Google Patents

Description

In order to reduce the risk of developing lifestyle-related diseases, it is effective to have a balanced diet as well as moderate physical activity. The "National Health Promotion Campaign in the 21st Century (Health Japan 21)" formulated under the Health Promotion Act states that the amount of physical activity necessary to maintain and improve health is "based on the results of epidemiological studies that have looked at the relationship between physical activity and mortality rates, etc., it is considered ideal to ensure 10,000 steps per day." However, according to the "Reiwa 1st Year National Health and Nutrition Survey Report" (Non-Patent Document 1), the average number of steps taken per day for men and women aged 20-64 is 7,226 steps, which is about 3,000 steps short on average.

Quercetin is a type of flavonoid, and is found in many plants, such as onions, either as it is or as a glycoside. In addition, the physiological activities of quercetin are known to include antioxidant, anti-inflammatory, antitumor, and vasodilatory effects, as well as the inhibition of muscle fibrosis (Patent Document 1), the inhibition of differentiation of satellite muscle cells from myofibroblasts to adipocyte-like cells (Patent Document 2), and the inhibition of muscle fibrosis (Patent Document 3).

In addition, Non-Patent Document 2 discloses that subjects were given a test beverage containing 275 mg of Sophora japonica polyphenols (enzymatically modified isoquercitrin) (the amount of quercetin or its glycoside is 71.6 mg in terms of quercetin, as described below) per day for seven days, and on the seventh day, they were made to exercise briefly on a bicycle ergometer. During high-intensity exercise of 70-90 watts, the amount of fat burned was significantly higher than that of subjects who had taken a control beverage that did not contain Sophora japonica polyphenols, but there was no significant difference in the amount of fat burned at exercise loads of less than 70 watts. According to the website of the bicycle ergometer manufacturer, exercise at 70 watts on the same bicycle is equivalent to approximately 6.5 METs at the exercise intensity described below (Non-Patent Document 3).

International Publication No. WO 2015/166887 International Publication No. 2016/175136 International Publication No. 2018/164221

"Reiwa 1st Year National Health and Nutrition Survey Report," Ministry of Health, Labour and Welfare, December 2020 Pharmacology and Therapy, 2008, Vol. 36, No. 10, pp. 919-930 KONAMI SPORTS CLUB Online Shop, "About the exercise effect of Aero Bike R", [online], [Searched on July 8, 2022], Internet <URL: https://online.konamisportsclub.jp/page/ksc/aboutaerobike/about_aerobike.html>

If the health-promoting effects of exercise can be enhanced by combining the intake of certain foods with exercise, people will be able to feel the benefits of exercise more, which will increase their motivation to exercise and ultimately lead to improved health. In particular, if the effects of low- to moderate-intensity exercise that can be continued on a daily basis could be enhanced, this would be ideal as it would contribute to the health of a wide range of people, regardless of age or exercise experience.

The present invention aims to provide a composition that contains as an active ingredient a component that can be safely ingested over a long period of time, and that enhances the body fat reduction effect of low- to medium-intensity exercise that can be easily continued on a daily basis.

The inventors have found that quercetin or its glycoside functions as an active ingredient that enhances the body fat reducing effect of low to moderate intensity exercise that can be continued on a daily basis (e.g., exercise with an intensity of 2.5 to 6.5 METs). In other words, the inventors have found that quercetin or its glycoside is suitable for reducing body fat when used in combination with low to moderate intensity exercise that can be continued on a daily basis (e.g., exercise with an intensity of 3 to 6.5 METs), and have thus completed the present invention.

That is, the present invention encompasses, but is not limited to, the following compositions for enhancing the effect of reducing body fat through exercise, methods for enhancing the reduction of body fat through exercise in a subject, and the like.
[1] A composition for enhancing the body fat reducing effect of exercise at an intensity of 2.5 to 6.5 METs, comprising quercetin or a glycoside thereof as an active ingredient.
[2] A composition for reducing body fat in a subject in combination with exercise, comprising quercetin or a glycoside thereof, wherein the intensity of the exercise is 2.5 to 6.5 METs.
[3] The composition according to [1] or [2], containing quercetin or its glycoside in an amount such that the total intake amount of quercetin or its glycoside in an adult human is 30 to 250 mg per day, calculated as a quercetin equivalent.
[4] The composition described in [1] or [2], which is a food or beverage.
[5] A method for enhancing reduction in body fat induced by exercise at an intensity of 2.5 to 6.5 METs in a subject, comprising the steps of: having a subject exercise at an intensity of 2.5 to 6.5 METs; and having the subject ingest a composition containing quercetin or a glycoside thereof (however, medical procedures on humans are excluded).
[6] A method for enhancing body fat reduction induced by exercise in a subject who exercises at a level of 2.5 to 6.5 METs three or more days per week, comprising having the subject ingest, at least three days per week, a composition containing quercetin or a glycoside thereof, wherein the composition contains quercetin or a glycoside thereof in an amount corresponding to a daily intake of 30 to 250 mg in terms of quercetin (however, medical procedures on humans are excluded).
[7] A method for enhancing body fat reduction induced by exercise in a subject who exercises at a level of 2.5 to 6.5 METs at least three days per week, comprising having the subject ingest a composition containing quercetin or a glycoside thereof at least three days per week, wherein the subject's intake of quercetin or a glycoside thereof is 30 to 250 mg per day in terms of quercetin (however, medical procedures on humans are excluded).
[8] A method for enhancing reduction of body fat caused by walking in a subject who walks 8,500 steps or more per day, comprising having the subject ingest a composition containing quercetin or a glycoside thereof, wherein the subject's intake of quercetin or a glycoside thereof is 30 to 250 mg per day in terms of quercetin (however, medical procedures on humans are excluded).
[9] A method for enhancing reduction in body fat caused by walking in a subject, the method comprising having the subject ingest a composition containing quercetin or a glycoside thereof at least three days a week for at least two weeks, the subject walking at least 8,500 steps per day at least three days a week for at least two weeks, the composition comprising quercetin or a glycoside thereof at least three days a week for at least two weeks, the amount of quercetin or a glycoside thereof ingested by the subject being 30 to 250 mg per day in terms of quercetin (however, medical procedures on humans are excluded).

The composition of the present invention contains as an active ingredient a component that can be safely ingested over a long period of time, and can enhance the effect of reducing body fat through low- to medium-intensity exercise. The present invention can promote reduction of body fat in subjects who perform low- to medium-intensity exercise in addition to their daily activities, thereby improving the subjects' motivation to exercise and contributing to the promotion of their health.

1 is a graph comparing the amount of change in abdominal total fat area at 12 weeks after the start of the study relative to the start of the study in each group in the study of Example 1 and the study of Comparative Example 1. Bars represent standard error. 1 is a graph comparing the change in abdominal subcutaneous fat area at 12 weeks after the start of the study relative to the start of the study in each group in the study of Example 1 and the study of Comparative Example 1. Bars represent standard error.

[Composition for enhancing the effect of low to moderate intensity exercise in reducing body fat]
The present invention includes a composition for enhancing the body fat reducing effect of low to moderate intensity exercise, which contains quercetin or a glycoside thereof as an active ingredient.

(Quercetin or its glycoside)
In this specification, "quercetin" refers to quercetin, a compound that is also called vitamin P and belongs to the flavonol group, a type of polyphenol. Quercetin is a compound represented by the following formula (I).

In the present invention, the term "quercetin glycoside" refers to a glycoside of the above-mentioned quercetin. The quercetin glycoside is a compound represented by the following general formula (II). In the following general formula (II), (X) _n represents a sugar chain, and n is an integer of 1 or more.

The sugar constituting the sugar chain represented by X that is glycosidically bonded to quercetin is, for example, glucose, rhamnose, galactose, glucuronic acid, etc., and is preferably glucose or rhamnose. Furthermore, n is not particularly limited as long as it is 1 or more, but is preferably 1 to 16, more preferably 1 to 8. When n is 2 or more, the X moiety may be composed of one type of sugar or may be composed of multiple types of sugars. In other words, when n is 2 or more, (X) _n may be a sugar chain composed of one type of sugar or may be a sugar chain composed of multiple types of sugars.

The above quercetin glycosides also include existing quercetin glycosides that have been treated with enzymes or the like to undergo glycosylation. The quercetin glycosides referred to in the present invention specifically include rutin, enzyme-treated rutin, quercitrin, isoquercitrin, enzyme-treated isoquercitrin, etc.

Rutin is a compound represented by the following formula (III):

In the present invention, it is particularly preferable to use an enzyme-treated product of rutin as the quercetin glycoside. Preferred examples of enzyme-treated products of rutin include rutin enzymatic hydrolysis products mainly composed of isoquercitrin obtained by enzymatically treating rutin to remove the rhamnose sugar chain portion, enzyme-treated rutin mainly composed of α-glucosylrutin obtained by enzymatically treating rutin to add one glucose molecule to the rutinose of rutin, and enzyme-treated isoquercitrin mainly composed of α-glucosylisoquercitrin obtained by treating rutin enzymatic hydrolysis products or isoquercitrin with a glycosyltransferase to bind a sugar chain consisting of 1 to 7 glucose units. Of these, it is preferable to use enzyme-treated isoquercitrin, which is a mixture of α-glucosylisoquercitrin having a sugar chain consisting of 1 to 7 glucose units.

Ingested quercetin glycoside is absorbed into the body through the digestive tract, and then converted into quercetin by the action of digestive or metabolic enzymes, and can exert the same effects as quercetin in the body.
In the present invention, only one type of quercetin or its glycoside may be used, or multiple types of compounds may be used. When multiple types of compounds are used, for example, quercetin and one or more types of quercetin glycosides may be used, or two or more types of quercetin glycosides may be used.

There are no particular limitations on the origin or manufacturing method for obtaining quercetin or its glycosides used in the present invention. For example, plants that are known to contain a lot of quercetin or its glycosides include buckwheat, sophora japonica, capers, apples, tea, onions, grapes, broccoli, mulukhiyah, raspberries, cowberries, cranberries, opuntia, leafy vegetables, and citrus fruits, and quercetin or its glycosides can be obtained from these plants.

(effect/use)
As will be shown in the Examples below, the composition of the present invention, when used in combination with low to medium intensity exercise, preferably low intensity exercise (hereinafter, also simply referred to as "the exercise"), has the effect of enhancing the body fat reducing effect of the exercise. Even in exercise of an intensity at which the effects of quercetin or its glycoside on muscle mass and basal metabolic rate are not yet evident, it is possible to significantly reduce body fat by taking quercetin or its glycoside. Although the details of the mechanism are unknown and do not limit the present invention, it is presumed that quercetin or its glycoside has the effect of increasing the amount of fat decomposition during the exercise.

Therefore, quercetin or its glycosides can be used in combination with the above exercise, and can be used to enhance the body fat reducing effect of the above exercise. Also, it can be used to enhance the body fat reducing effect when the above exercise is performed in addition to physical activities in daily life or work (collectively referred to as daily activities). Also, it can be used to enhance the body fat reducing effect when the above exercise is performed regularly and continuously in addition to daily activities. Here, the use to enhance the body fat reducing effect of the above exercise includes not only the promotion or increase of the reduction in body fat by the above exercise, but also, for example, the promotion or increase of the breakdown or burning of body fat by the above exercise, which results in an enhancement of the body fat reducing effect by the above exercise.

The exercise is 6.5 METs or less, preferably 6.2 METs or less, more preferably 6.0 METs or less, even more preferably 5.5 METs or less, even more preferably 5.0 METs or less, even more preferably 4.5 METs or less, and particularly preferably 4.0 METs or less, so that it can be continued on a daily basis and still enjoy the body fat reduction enhancing effect of the present invention. The exercise is, for example, 2.5 METs or more, preferably 3.0 METs or more, more preferably 3.2 METs or more, and more preferably 3.5 METs or more. Here, METs (Metabolic Equivalent of Task) is an index that indicates the intensity of various physical activities, and indicates the energy consumption of each physical activity when the energy consumption at rest is set to 1.0.

Low to moderate intensity exercise (particularly low intensity exercise) is an exercise that can be continued on a daily basis, and by continuing to do so, the body fat reduction enhancing effect of the present invention is further exerted. In particular, the composition of the present invention has the effect of significantly promoting body fat reduction in subjects who regularly and continuously perform low to moderate intensity exercise (particularly low intensity exercise) in addition to their daily activities.

In this specification, the number of METs for each physical activity related to the above exercises can be, for example, the values listed in the "Revised Physical Activity METs Table" (revised on April 11, 2012, URL: https://www.nibiohn.go.jp/files/2011mets.pdf) created by the National Institute of Health and Nutrition.

Examples of the above exercises include walking (normal walking, brisk walking, walking, etc.), running (jogging, running, etc.), exercises (aerobics, radio calisthenics, yoga, pilates, tai chi, etc.), dancing, cycling, climbing and descending stairs, strength training, carrying luggage, climbing and descending stairs, strength training, various sports (soccer, baseball, golf, basketball, badminton, swimming, tennis, softball, volleyball, table tennis, rugby, surfing, skiing, etc.), as well as low- to medium-intensity exercise within the above-mentioned MET number range among physical activities in daily life or work (housework, cooking, tidying up, cleaning, etc.). It is preferable to perform these exercises in addition to daily activities.

The duration of the above exercise can be set appropriately depending on the subject's physique, age, and physical condition, the type and intensity of exercise, the target amount of body fat reduction, etc., and can be, for example, 3 minutes or more, 5 minutes or more, 10 minutes or more, 30 minutes or more, 60 minutes or more, or 2 hours or more, or, for example, 10 hours or less, or 5 hours or less.

The total time for the above exercise per day can be set appropriately depending on the subject's physique, age, and physical condition, the type and intensity of exercise, the target amount of body fat reduction, etc., but is, for example, at least 10 minutes, at least 30 minutes, at least 60 minutes, at least 2 hours, or at least 5 hours, and is, for example, at most 10 hours.

From the viewpoint of more prominently achieving the effects of the present invention, it is preferable that the above exercise is performed repeatedly (e.g., periodically). The frequency can be set appropriately depending on the type and intensity of the exercise, the target amount of body fat reduction, etc., and is, for example, every day, 1 to 6 times a week, or 1 to 3 times a week. Alternatively, the frequency of exercise is at least 1 day a week, preferably at least 2 days a week, more preferably at least 3 days a week, even more preferably at least 4 days a week, even more preferably at least 5 days a week, even more preferably at least 6 days a week, and most preferably every day. In addition, from the viewpoint of more prominently achieving the effects of the present invention, the period for which the above exercise is performed repeatedly is preferably at least 2 weeks, more preferably at least 4 weeks, even more preferably at least 8 weeks, and even more preferably at least 12 weeks.

When the exercise is walking, the suitable amount of exercise per day is preferably 8,500 steps or more, more preferably 9,000 steps or more, and even more preferably 10,000 steps or more in total steps. According to the Ministry of Health, Labor and Welfare's "Reiwa 1st Year National Health and Nutrition Survey Report," the average number of steps per day for men and women aged 20-64 is 7,226 steps. Considering this, when the exercise is walking consciously performed in addition to daily activities, the amount of exercise per day is preferably 1,000 steps or more, more preferably 2,000 steps or more, and even more preferably 3,000 steps or more. The exercise intensity of normal walking varies depending on the walking speed and walking environment, but is approximately 2.5 to 4.0 METs.

The exercise to be used in combination with the composition of the present invention is preferably about 1.0 to 3.0 exercises per day, more preferably 1.0 to 2.5 exercises, and even more preferably 1.0 to 2.0 exercises, expressed in "exercise (Ex)" (METs-hours), which is a unit obtained by multiplying METs, which are the exercise intensity, by the exercise time (hours). These exercises may be added to daily activities.

Specific examples of low to moderate intensity exercise to be used in conjunction with the compositions of the present invention include, but are not limited to, the following:
- Walking at least 8,500 steps per day, more preferably at least 9,000 steps, and even more preferably at least 10,000 steps.
- Consciously walking 1,000 steps or more, more preferably 2,000 steps or more, and even more preferably 3,000 steps or more in addition to your daily activities.
- Walking for at least 10 minutes, more preferably at least 15 minutes, more preferably at least 20 minutes, more preferably at least 25 minutes, and even more preferably at least 30 minutes, consciously performed in addition to daily activities.
- Cycling for at least 5 minutes, more preferably at least 10 minutes, and even more preferably at least 15 minutes.
- NHK Radio Exercise 1 or 2 once each.
・One game of bowling.
・One game of table tennis.
- Playing video games using the whole body for 10 minutes or more, preferably 15 minutes or more, more preferably 20 minutes or more, more preferably 30 minutes or more.
- Housework, cooking, tidying up, and cleaning for more than 10 minutes, preferably more than 15 minutes, more preferably more than 20 minutes, and more preferably more than 30 minutes.

The timing of ingestion of the composition of the present invention is not particularly limited in relation to the timing of the exercise, and the composition may be ingested when not exercising, or may be ingested before, during, or after the exercise.

Specific examples of body fat that can be reduced by using the composition of the present invention in combination with the above-mentioned exercise include total fat, subcutaneous fat, and visceral fat, and the reduction effect is particularly remarkable for total fat and subcutaneous fat. An example of the part of the body fat that can be measured is the abdomen. The reduction effect of these body fats can be confirmed, for example, by the method for measuring abdominal fat area described in the Examples.

(Form of composition)
The composition of the present invention can be provided in the form of a drug, for example, but is not limited to this form. The drug can be provided as a composition as it is, or as a composition containing the drug.

The composition of the present invention can be provided in the form of, for example, a food or drink, a medicine, a quasi-drug, feed, etc., but is not limited thereto. The composition of the present invention may itself be a food or drink, a medicine, a quasi-drug, feed, etc., or may be a preparation or material such as an additive used therein. In one embodiment, the composition of the present invention is preferably an oral composition.

In one embodiment, the composition of the present invention is preferably a food or beverage, a medicine (preferably an oral medicine) or a quasi-drug, more preferably a food or beverage or an oral medicine, even more preferably a food or beverage, and even more preferably a beverage.

The above-mentioned foods and beverages are not particularly limited, and examples thereof include general foods and beverages, health foods, functional foods, foods for specified health uses, nutritional functional foods, health foods, dietary supplements, supplements, foods for the sick, or food additives, or raw materials thereof. The form of the above-mentioned foods and beverages is also not particularly limited, and in addition to the usual forms of beverages or foods, they can also be in various formulation forms such as oral solid preparations such as powders, tablets, coated tablets, fine granules, granules, powders, pills, capsules, dry syrups, and chewable preparations; and oral liquid preparations such as oral liquid preparations and syrups.

Examples of the above beverages include tea beverages (green tea, oolong tea, roasted green tea, jasmine tea, barley tea, brown rice tea, buckwheat tea, rooibos tea, mate tea, etc.), coffee beverages, flavored water beverages, sports drinks, carbonated drinks, vegetable and fruit beverages (green juice, vegetable juice, fruit juice, etc.), and alcoholic beverages (beverages with an alcohol content of 1% or more), but tea beverages, flavored water beverages, sports drinks, or carbonated drinks are preferred, and tea beverages are more preferred.

As long as the beverage is consumed by the subject, there are no particular limitations on the form in which it is distributed on the market or the blending ratio of each ingredient contained therein. Therefore, the beverage may be a concentrate such as a powder or syrup, and also includes beverages that are diluted, dissolved, or infused (with tea leaves or tea bags, etc.) with water, alcohol, carbonated water, etc. when consumed.

For beverage products that are to be consumed after dilution or the like, the concentration or amount of each component of the beverage described in this specification represents the concentration or amount when consumed.
The dosage form of the above-mentioned medicine or quasi-drug may be a dosage form suitable for the administration form, and may be a medicine for human use or a medicine for non-human animals. The administration form of the medicine or quasi-drug may be oral administration or parenteral administration, but oral administration is preferred.

Dosage forms of oral medicines include, for example, oral solid preparations such as tablets, coated tablets, fine granules, granules, powders, pills, capsules, dry syrups, and chewable preparations; and oral liquid preparations such as oral liquid preparations and syrups. Dosage forms of parenteral medicines include, for example, injections, infusions, topical preparations, suppositories, and transdermal preparations.

Examples of the above feed include livestock feed for cows, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, rats, mice, etc.; and pet food for dogs, cats, small birds, etc.

The composition of the present invention may contain any additives or components other than the active ingredient quercetin or its glycoside, as long as the effects of the present invention are not impaired. As these additives and components, those that can generally be used in foods and beverages, medicines, quasi-drugs, feed, cosmetics, etc., can be used. Examples of optional additives or components include physiologically active ingredients such as vitamins such as vitamin E and vitamin C, minerals, and nutritional ingredients, as well as excipients, binders, emulsifiers, tonicity agents (isotonicity agents), buffers, solubilizers, preservatives, stabilizers, antioxidants, colorants, coagulants, coating agents, fragrances, etc., which are blended in formulations. These may be used alone or in combination of two or more.

In addition to the above, ingredients such as materials used in foods and beverages, medicines, quasi-drugs, feed, cosmetics, etc. can be appropriately blended depending on the application.
The composition of the present invention may be labeled on its packaging, container, or instruction manual with one or more information such as intended use, type of active ingredient, the above-mentioned effects, and method of use (e.g., method of ingestion, method of administration).

As such indications, the present invention may be provided with at least one indication selected from, for example, (1) an indication that the composition has the effect of enhancing the body fat reduction effect of the exercise, promoting or increasing the reduction of body fat by the exercise, promoting or increasing the breakdown of body fat by the exercise, or promoting or increasing the burning of body fat by the exercise, (2) an indication that the composition can be used to obtain the effect (for example, indications recommending the combined use of the composition of the present invention and the exercise, such as "drink and walk," "combine with low to medium intensity exercise," or "add to your daily exercise habits"); and (3) an indication of subjects who require or desire the effect (for example, "for those who want to reduce body fat while exercising daily," "for those who want to reduce body fat with light exercise," "for those who want to reduce body fat by walking," etc.).

The content of quercetin or its glycoside in the composition of the present invention is not particularly limited and can be appropriately set depending on the form, etc. For example, the total content of quercetin or its glycoside in the composition of the present invention (total content of quercetin and its glycoside) is, in terms of quercetin relative to the total amount of the composition, for example, 0.001% by weight or more, preferably 0.002% by weight or more, more preferably 0.003% by weight or more, and for example, 99.9% by weight or less, preferably 95% by weight or less, more preferably 80% by weight or less, and even more preferably 45% by weight or less.

When the composition of the present invention is a beverage, the total content of quercetin or its glycosides in the composition is, in terms of quercetin, preferably 0.002 to 0.5% by mass, more preferably 0.003 to 0.1% by mass, more preferably 0.005 to 0.05% by mass, and even more preferably 0.01 to 0.02% by mass, based on the total amount of the composition.

When referring to the concentration or amount of a beverage ingredient in this specification, it refers to the concentration or amount in the final product, unless otherwise specified. The ingredient may decompose to some extent during processes such as sterilization, and the amount to be added to the beverage can be determined taking into account the amount of decomposition, but usually the concentration or amount at the time of addition will be roughly the same as that in the final product.

When the composition of the present invention is a supplement, the total content of quercetin or its glycosides in the composition can be, in terms of quercetin, preferably 0.1 to 50% by mass, more preferably 0.2 to 20% by mass, and even more preferably 0.3 to 10% by mass, based on the total amount of the composition.

The amount of quercetin glycoside is measured by the following method, except where otherwise specified, using QG1 to QG7 as the participating components: that is, using quercetin 3-O-glucoside (QG1) as the standard substance, a calibration curve is created by using high performance liquid chromatography (HPLC) and the area of ultraviolet absorbance at 350 nm and the concentration of the standard substance. Since quercetin glycoside is hydrolyzed to quercetin in the small intestine, QG1 to QG7 are considered to be equivalent in terms of physiological activity, and the 3-position glycoside of quercetin all have a maximum absorption at 350 nm regardless of the length of the sugar chain, and the absorbance depends on the aglycone part, quercetin. Therefore, although the molecular weights are different, QG1 to QG7 are considered to be equivalent in terms of molar absorbance, and the participating components are quantified in terms of QG1. Specifically, the analytical sample is subjected to HPLC under the same conditions as the standard substance, and a peak that coincides with the elution retention time of the standard substance is identified in the obtained chart. Then, the peaks of quercetin glycosides QG2 to QG7 detected before the QG1 peak are identified (if any), and the quercetin glycoside content in the analytical sample is calculated from the sum of the areas of each peak using a calibration curve created using a standard substance.

From another perspective, when the composition is a bottled beverage having a volume of 350 to 500 ml, the amount of quercetin or its glycoside, converted to the QG1 below, per bottle of beverage can be 10 to 1800 mg, preferably 50 to 900 mg, more preferably 100 to 500 mg, and even more preferably 100 to 250 mg.

In this specification, unless otherwise specified, the amount of quercetin in a beverage refers to the amount of quercetin produced by hydrolysis of QG1, calculated as the total amount of quercetin glycosides, converted into QG1 (e.g., isoquercitrin). The amount of quercetin produced by hydrolysis of QG1 can be calculated by (amount of quercetin glycoside/464) x 302, where quercetin has a molecular weight of 302 and QG1 has a molecular weight of 464.

(Dosage/Dosage)
The composition of the present invention can be ingested or administered in a suitable manner depending on the form of the composition. The composition of the present invention may be administered or ingested orally, or may be administered parenterally in the form of an injection or the like, but is preferably administered or ingested orally.

The intake amount (which can also be called the dosage amount) of the composition of the present invention is not particularly limited, and may be set appropriately depending on the dosage form, administration method, etc. In one embodiment, for example, when the composition of the present invention is administered or ingested orally or parenterally to a human (adult) subject, the intake amount of the composition of the present invention is, in terms of the total intake amount of quercetin or its glycosides (total intake amount of quercetin and its glycosides), preferably 0.1 mg to 8000 mg per day in quercetin equivalent, more preferably 0.3 mg to 4000 mg, even more preferably 1.0 mg to 1000 mg, even more preferably 10 mg to 500 mg, and particularly preferably 30 mg to 250 mg. It is preferable to orally administer or ingest the above amount, for example, 1 to 3 times a day.

The intake period and number of intakes of the composition of the present invention are not particularly limited. For example, the above amount may be taken in divided doses, for example, 1 to 3 times a day, on the day of low to medium intensity exercise, or may be taken before or after the day of exercise, such as the day before or the day after exercise. From the viewpoint of more prominently achieving the effects of the present invention, it is preferable to take the composition of the present invention daily, for example, 1 to 6 times a week, or 1 to 3 times a week, regardless of whether exercise is performed or not. From the viewpoint of more prominently achieving the effects of the present invention, it is preferable to take the composition of the present invention for a period of 2 weeks or more, 4 weeks or more, 8 weeks or more, or 12 weeks or more. For example, during such a period, it is preferable to take the composition of the present invention in the above amount every day, 1 to 6 days a week, or 1 to 3 days a week, and to repeatedly (for example, periodically) perform the low to medium intensity exercise described above during the same period.

(subject)
The subject to which the composition of the present invention is administered or ingested (hereinafter, also simply referred to as the administration subject) is preferably an animal, more preferably a mammal (human and non-human mammal), and even more preferably a human. Examples of non-human mammals include cows, horses, goats, dogs, cats, rabbits, mice, rats, guinea pigs, monkeys, etc. In addition, the administration subject of the present invention is preferably a subject who needs or desires to enhance the effect of reducing body fat by low to medium intensity exercise. For example, suitable subjects include subjects who do not exercise enough on a daily basis or who have limited time available for exercise, or subjects whose motor function has decreased due to weight gain, aging, etc.

In addition, individuals with a BMI (Body Mass Index) of 22 or more (preferably 25 or more, for example 25 or more and less than 30 (obesity (grade I))) are also suitable subjects for the composition of the present invention. For these subjects, the composition of the present invention is preferable because low to medium intensity exercise is preferable to reduce the burden on the body, and the effects of the present invention are more pronounced compared to normal weight individuals with a BMI of 18.5 to less than 25.

[Method of enhancing body fat reduction through low- to moderate-intensity exercise]
The present invention includes a method for enhancing body fat reduction induced by low- to moderate-intensity exercise in a subject, comprising the steps of having the subject perform low- to moderate-intensity exercise (e.g., exercise with an intensity of 2.5 to 6.5 METs) and having the subject ingest a composition containing quercetin or a glycoside thereof.

The above method may be a therapeutic method or a non-therapeutic method, the term "non-therapeutic" being used herein to mean a method that does not include medical procedures, i.e. surgery, treatment or diagnosis.
The dosage of quercetin or its glycoside may be an amount that enhances the reduction of body fat caused by exercise in a subject, i.e., an effective amount, and is not particularly limited. For example, it is preferable to administer or ingest the dosage described in the above section [Composition for enhancing the effect of reducing body fat caused by low- to medium-intensity exercise]. Quercetin or its glycoside may be administered or ingested as it is, or may be administered or ingested as a composition containing quercetin or its glycoside. For example, the composition of the present invention described above can be administered or ingested. Quercetin or its glycoside, low- to medium-intensity exercise, subjects to be administered, administration method, dosage, and preferred aspects thereof are the same as those in the composition of the present invention described above. According to the method of the present invention, the effect of reducing body fat caused by low- to medium-intensity exercise can be enhanced safely without causing side effects.

The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to these examples.
Example 1: Human study of the effects of combining quercetin glycoside with low- to moderate-intensity exercise
(Test Overview)
(1) Test Beverages The test beverage was 500 mL of green tea beverage containing 110 mg of quercetin glycoside as isoquercitrin (71.6 mg in terms of quercetin). The control beverage was 500 mL of green tea beverage without quercetin glycoside, which was prepared so as to be indistinguishable from the test beverage in terms of shape, color, flavor, etc. The nutritional composition analysis values of the test beverage and the control beverage were 0 kcal for energy, 0 g for protein, 0 g for fat, and 0 g for carbohydrate.

(2) Study subjects: Healthy men and women aged 20 to 65 years, with obesity level I (BMI of 25 kg/ ^m2 to 30 kg/ ^m2 ) were selected as study subjects. In addition, those who regularly engaged in strenuous exercise (jogging, aerobics, strength training, swimming, etc.), heavy alcohol drinkers and smokers, those who continuously took medicines, those who were pregnant or hoping to become pregnant during the study period, and those who were breastfeeding, those who had allergies to the test beverages, and others who the study investigator deemed inappropriate as study subjects were excluded.

(3) Test Method This test example was conducted as a randomized, double-blind, placebo-controlled, parallel group comparative study with a 12-week intake period. The test subjects were assigned to a test beverage group (n=74) and a control beverage group (n=74) to ensure randomness. The test subjects were instructed to continue to consume one test beverage or control beverage per day according to the allocation from the test conducted at the start of the intake of the test beverage until the day before the test 12 weeks after intake, and to walk about 3000 steps (target range: 3000±500 steps) more per day than the average number of steps per week of each test subject measured before the test. In addition, during the test period, the subjects were instructed not to change their diet and exercise other than walking significantly from before the start of the test, and to keep them as constant as possible. The exercise intensity of walking is generally about 2.5 to 4.0 METs. In addition, 3000 steps generally corresponds to about 30 minutes of walking.

(4) Test Items The test subjects were examined for the following items. Test data was collected at the start of intake (week 0), and at 8 and 12 weeks after intake.

<Abdominal fat area>
Abdominal fat area was measured according to the method of Tokunaga et al. (Tokunaga K et al. Int. J. Obes., 1983, Vol.7, pp.437-45) using a computer tomography (CT) scanner system (Hitachi whole body X-ray device ROBUSTO Ei, Hitachi Medical Corporation, Tokyo, Japan). The measurement site was the umbilicus (if the kidney and ilium were included in the image taken at the position of the umbilicus, the image was taken again with the fourth lumbar bone as the center), and the image was taken at the end-expiratory phase. The CT imaging conditions were tube voltage: 120 kVp, tube current: AEC function, slice thickness: 5 mm, and film processing was window level "0" and window width "1000". The obtained images were analyzed using visceral fat measurement PC software (Fat Scan Premium, East Japan Technical Research Institute, Ibaraki Prefecture, Japan).

<Steps>
The test subjects were asked to record the number of steps they took each day in a diary using a pedometer, and the average number of steps taken per day was calculated for each period from the start of intake (week 0) through the 12-week test period after intake.

(5) Statistical Analysis The group of all randomized test subjects excluding test subjects who did not consume any test beverages and test subjects with no post-allocation data was defined as the Full Analysis Set (FAS), and the group of FAS that complied with the study implementation protocol was defined as the Per Protocol Set (PPS). Analysis was performed on the PPS.

Abdominal fat area was evaluated using a linear mixed effects model with group, test time, and group-test time interaction as fixed effects, and test subject as a random effect. Comparisons were made between groups consuming each test beverage at each test time and with the start of intake (week 0). Comparisons between intake groups were evaluated using an F test to test for homogeneity of variance, with Student's t-test used in the case of homogeneity of variance and Welch's t-test used in the case of unequal variance. Comparisons with the start of intake (week 0) were evaluated using a paired t-test. The statistical significance level was 5%, and the test was two-sided. SAS software (R9.4, SAS Institute Japan), SPSS (SPSS Statistics ver. 27, IBM), etc. were used for compilation and analysis.

(result)
(1) Analysis subjects At the time of allocation, there were 148 subjects, of which 143 subjects (73 in the test beverage group and 70 in the control beverage group) started taking the beverage (FAS). Background data of the test subjects at the time of allocation is shown in Table 1. The number of test subjects related to the PPS is listed in each table of results.

(2) Abdominal total fat area The results are shown in Tables 2 and 3. A significant reduction in change in abdominal total fat area was observed with 12 weeks of continuous ingestion of the test beverage compared to ingestion of the control beverage (interaction between group and test time by linear mixed effects model: p<0.05, main effect of group: p<0.05). Furthermore, a significant reduction in change in abdominal total fat area was observed in the test beverage group compared to the control beverage group at tests 8 and 12 weeks after ingestion (p<0.05). Furthermore, a significant reduction in abdominal total fat area was observed in the test beverage group compared to the start of ingestion (week 0) at tests 8 and 12 weeks after ingestion (p<0.05).

(3) Abdominal subcutaneous fat area The results are shown in Tables 2 and 3. A significant reduction in abdominal subcutaneous fat area change was observed with 12 weeks of continuous ingestion of the test beverage compared to ingestion of the control beverage (interaction between group and test time by linear mixed effects model: p<0.05, main effect of group: p<0.05). The test beverage group showed a significant reduction in abdominal subcutaneous fat area change at tests 8 and 12 weeks after ingestion compared to the control beverage group (p<0.05). Furthermore, the test beverage group showed a significant reduction in abdominal subcutaneous fat area at tests 8 and 12 weeks after ingestion compared to the start of ingestion (week 0) (p<0.05).

(4) Number of steps The average number of steps per day before ingestion was 6,890.2 steps/day for the test beverage group and 7,000.4 steps/day for the control beverage group. The average number of steps per day during the ingestion period (from the day ingestion began until the examination 12 weeks after ingestion) was 9,813.0 steps/day for the test beverage group and 10,000.0 steps/day for the control beverage group, showing roughly similar trends over time for both groups.

Comparative Example 1: Human test of the effect of quercetin glycoside without exercise
(Test Overview)
A randomized, double-blind, placebo-controlled study was conducted on subjects aged 20-65 (81 men, 119 women) with a BMI between 25 and 30. Subjects were randomly divided into two groups and asked to consume a beverage containing 0 mg or 110 mg of quercetin for 12 weeks (89 subjects in the control group (36 men, 53 women) and 83 subjects in the test group (33 men, 50 women)). Details of the method and results of this study are as described in the paper by Egawa et al. (Egawa et al., Pharmacology and Treatment, 2012, Vol. 40, No. 6, pp. 495-503).

In other words, the test in Comparative Example 1 differs from the test in Example 1 above in that no exercise was instructed, but it was conducted under the same conditions in terms of beverage, subjects (adult males and females with obesity level I), region, and season.

(result)
Compared to the start of the study, total abdominal fat area 12 weeks after the start of the study was -5.32±2.42 ^cm2 in the test beverage group and +4.97±2.34 ^cm2 in the control beverage group (both values are standard error). The change in total abdominal fat area in the test beverage group compared to the control beverage group (examination 12 weeks after ingestion) was -10.30 ^cm2 .

Compared to the start of the study, abdominal subcutaneous fat area 12 weeks after the start of the study was -2.31±1.73 ^cm2 in the test beverage group and +0.64±1.56 ^cm2 in the control beverage group. The change in abdominal subcutaneous fat area in the test beverage group compared to the control beverage group (examination 12 weeks after ingestion) was -2.94 ^cm2 .

[Considerations based on a comparison between Example 1 and Comparative Example 1]
Graphs comparing the change in abdominal total fat area and abdominal subcutaneous fat area from the start of the study 12 weeks after the start of each study are shown in Figures 1 and 2. By combining intake of quercetin glycoside with low- to medium-intensity exercise, such as walking about 3,000 steps, abdominal total fat area and abdominal subcutaneous fat area were significantly reduced compared to the case of no exercise. Such an effect of combining intake of quercetin glycoside with low- to medium-intensity exercise was not observed in the group that consumed a control drink that did not contain quercetin glycoside.

These results suggest that quercetin glycoside significantly enhances the body fat reduction effect of low- to moderate-intensity exercise.

Claims (2)

A method for enhancing reduction in body fat caused by walking in a subject who continuously walks at least 8,500 steps per day with an exercise intensity of 2.5 to 4.0 METs for at least 8 weeks, comprising having the subject ingest a composition containing quercetin or a glycoside thereof, wherein the subject's intake of quercetin or a glycoside thereof is 30 to 250 mg per day in terms of quercetin (however, medical procedures on humans are excluded). A method for enhancing reduction in body fat caused by walking in a subject, the method comprising having a subject who walks at least 8,500 steps per day with an exercise intensity of 2.5 to 4.0 METs , at least 3 days per week for at least 8 weeks , ingest a composition containing quercetin or a glycoside thereof at least 3 days per week for at least 8 weeks, the amount of quercetin or a glycoside thereof ingested by the subject being 30 to 250 mg per day in terms of quercetin (however, medical procedures on humans are excluded).