Energy metabolism promoting composition, energy metabolism promoter, and method for activating adipose tissue

Abstract

Provided are: an energy metabolism promoting composition and an energy metabolism promoter effective for improving obesity conditions and preventing obesity; and a method for activating an adipose tissue using the same. The energy metabolism promoting composition contains trigonelline as an active ingredient. The composition may contain coffee-derived trigonelline as an active ingredient. The composition may further contain caffeine, or may further contain chlorogenic acids. The composition may further contain a lipid. The trigonelline may be extracted from raw coffee beans or roasted coffee beans. The energy metabolism promoter comprises any one of the above-mentioned energy metabolism promoting compositions. The energy metabolism promoter may be in the form of a beverage, food, a supplement or a medicine. The method for activating an adipose tissue uses any one of the above-mentioned energy metabolism promoting compositions.

Description

Energy metabolism promoting composition, energy metabolism promoting agent, and method for activating adipose tissue

[0001] The present invention relates to an energy metabolism promoting composition and an energy metabolism promoting agent that are effective in improving obesity and preventing obesity, and a method for activating adipose tissue using them.

[0002] Coffee is one of the most widely consumed beverages in the world and is a popular beverage rich in functional components, such as caffeine and chlorogenic acids. Previous epidemiological studies have suggested that coffee consumption is effective in preventing chronic diseases such as type 2 diabetes, Parkinson's disease, and liver disease. It has also been reported that individuals who drink three to four cups of coffee per day have a 24% lower risk of death compared to non-drinkers. For example, a cross-sectional cohort study using the US National Health and Nutrition Examination Survey showed that higher coffee intake was associated with a dose-response relationship between total body fat and trunk body fat percentage in US women. In recent years, the beneficial physiological effects of trigonelline (1-methylpyridin-1-ium-3-carboxylate), a component found in coffee beans, have attracted attention. Trigonelline is found in the highest concentration in green coffee beans and is an alkaloid with various functional properties, including hypoglycemic, hypolipidemic, and cognitive improvement effects. Trigonelline is a component derived from nicotinic acid and is found in high concentrations not only in green coffee beans but also in some legumes. Non-Patent Document 1 discloses that trigonelline's favorable physiological effects include its effectiveness in protecting the heart and liver and treating hyperglycemia, hypercholesterolemia, neurological and hormonal disorders, and cancer. Sakurajima radish is also found to contain high concentrations of trigonelline, and its continuous intake as edible radish has been described to improve vascular endothelial function (see Non-Patent Document 2).

[0003] Trigonelline is also found in various plants, and is ingested in trace amounts on a daily basis without even realizing it. However, it is heat-sensitive and converts to nicotinic acid when heated, resulting in the loss of its effects. Patent Documents 1 and 2 disclose methods for producing functional foods containing trigonelline, which are expected to prevent brain aging and Alzheimer's disease by suppressing trigonelline degradation through adjusting roasting conditions during trigonelline extraction from coffee beans. Patent Document 3 also discloses a composition containing trigonelline, which increases nicotinamide adenine dinucleotide (NAD+) levels in cells and tissues, thereby improving muscle fatigue and mobility. This document relates to a composition and method for producing intracellular NAD+ / NADH using trigonelline, but its examples mainly focus on NAD+, and no results are provided on adipocytes (including precursor cells) or animal white fat, brown fat (or beige fat) through cell or animal testing. That is, Patent Document 3 makes no mention of adipocytes, and does not disclose or suggest a composition that promotes energy metabolism using trigonelline, or a composition that promotes energy metabolism by promoting beige coloration of white adipose tissue. Furthermore, Patent Document 4 discloses a fat-burning composition that contains pipecolic acid and trigonelline as active ingredients and promotes fat burning in adipocytes. However, this document states that "the combined use of pipecolic acid and trigonelline can promote the expression of the UCP-1 gene" (paragraph 0011 of Patent Document 4), which indicates that the combined use of pipecolic acid and trigonelline promoted the expression of the UCP-1 gene. Furthermore, the court also states that "as a result, neither pipecolic acid (PIP) nor trigonelline (TRG) had any significant effect when used alone, but when pipecolic acid was adjusted to 1 μM and trigonelline to 10 μM, it was found that the relative gene expression level of UCP-1 increased significantly (1.58 compared to 1.00 in the control)" (paragraph 0047, Table 1), which shows that trigonelline alone without the use of pipecolic acid (PIP) showed almost no difference from the control.Unlike white fat cells, which store excess energy in the form of triglycerides, brown fat cells produce heat and promote energy consumption through the action of the UCP-1 gene present in the inner mitochondrial membrane. Therefore, an increase in the expression level of the UCP-1 gene does not necessarily promote the beige transformation of white fat cells.

[0004] A recently discovered favorable effect of trigonelline is reported in Non-Patent Document 3, which reports its effect on the beigeification of white adipocytes. Adipose tissue is classified into two types: tissue composed of white adipocytes (White Adipose Tissue: WAT), which accumulate excess energy as neutral fat and cause obesity, and tissue composed of brown adipocytes (Brown Adipose Tissue: BAT), which have the effect of preventing obesity by burning fat and generating heat. It has been considered problematic that in adults, tissue composed of brown adipocytes (BAT) shrinks or disappears significantly with age, making obesity more likely to progress. However, it has been revealed that beige adipocytes, which have the effect of burning fat similar to brown adipocytes, differentiate from white adipocytes in white adipocyte tissue when exposed to cold or other stimuli. Similar to brown adipocytes (BAT), beige adipocytes consume energy through non-shivering thermogenesis mediated by uncoupling protein 1 (UCP-1) and contribute to maintaining core body temperature in adults in cold environments. Thus, beige adipocytes, along with endogenous classical brown adipocytes, activate fat metabolism (thermoconversion), which is effective in preventing and improving obesity. Therefore, the beigeification of white adipocytes has attracted attention as a strategy for combating obesity.

[0005] Previous research has revealed that cold stimulation and exercise release noradrenaline from the sympathetic nerve terminals, which in turn stimulates β3-adrenergic receptors (β3ARs), promoting the beige transformation of white fat cells and the activation of brown fat cells. Furthermore, in addition to these physical stimuli, oral administration of capsaicin, the spicy component of chili peppers, and its analogous substance, capsinoid, has also been shown to activate the sympathetic nervous system, resulting in the beige transformation of white fat cells and the activation of brown fat cells.

[0006] In addition to capsaicin, it has been found that ω-3 polyunsaturated fatty acids such as EPA and DHA contained in fish oil, as well as various food components such as menthol, allicin, and catechin, also have the effect of activating brown fat cells via sympathetic nervous system activation. Expectations are high for the obesity-suppressing effects of these various food-derived components.

[0007] As shown in Non-Patent Document 3, in the presence of trigonelline, white adipocytes turn beige, and therefore trigonelline is expected to have favorable effects on preventing and improving obesity. However, these results are based solely on observations of the effects at the cellular level, and it is unclear whether trigonelline has any effect on improving fat metabolism and preventing obesity in actual biological systems, and it is unclear whether it is effective in animals or humans.

[0008] Selective β3-adrenergic receptor agonists such as mirabegron, CL316243, BRL37344, and others are already known, and like trigonelline, these are expected to be effective in converting white adipocytes to beige and activating brown adipocytes. However, these have similar chemical structures to endogenous neurotransmitters such as adrenaline and noradrenaline, and there are concerns that they may increase high blood pressure and heart rate. Therefore, they should be avoided in patients with cardiovascular complications such as arrhythmia, and are contraindicated in patients already receiving antiarrhythmic drugs. Therefore, it is not recommended to take these drugs on a daily basis to prevent or improve obesity.

[0009] There is a demand for energy metabolism-promoting compositions with these effects in the form of beverages or supplements that can be consumed daily to prevent or improve obesity, as well as methods for activating adipose tissue using such compositions. However, it is completely unclear whether such effects can actually be achieved by orally ingesting trigonelline or other ingredients.

[0010] Non-Patent Document 4 reports a risk assessment of trigonelline intake, finding that it is safe when taken daily in coffee drinks, etc., but no conclusion can be reached regarding the health effects of high-concentration intake due to a lack of data. Therefore, there is a need for a safe and effective energy metabolism-promoting composition that does not cause harmful side effects in humans, and a method for activating adipose tissue using the same.

[0011] In obese white adipose tissue, macrophage infiltration and activation induces a chronic inflammatory response, which is thought to be the main cause of the development of metabolic disorders associated with obesity. Because inflammatory cytokines act as mediators of the inflammatory response, suppressing this inflammatory response is an important strategy for preventing and improving obesity.

[0012] The progression of obesity is not just a problem of the increase in the size of lipid droplets inside white adipocytes, but also because white adipocytes themselves break down fat and supply energy sources as fatty acids to every corner of the body, and secrete various adipocytokines, which contribute to appetite suppression and improved insulin sensitivity. It is known that when white adipocytes accumulate lipid droplets and become enlarged, the secretion of adipocytokines decreases, and insulin resistance increases.

[0013] The nuclear transcription factor PPARγ, which acts on adipocyte differentiation and other lipid and glucose metabolism, is highly expressed in adipocyte tissue, and stimulating this factor is expected to enhance energy metabolism and be effective in preventing and improving obesity. Rosiglitazone, a selective agonist for PPARγ, is known as a thiazoline antidiabetic drug, and it is known that adding this to highly activate PPARγ promotes white adipocyte differentiation, reduces the size of white adipocytes, and promotes the secretion of adipocytokines. Furthermore, it is known that adding a PPARγ agonist also promotes the beige color of white adipocytes.

[0014] However, it has been pointed out that thiazolidinedione drugs such as rosiglitazone may cause cardiovascular disorders, and it is not preferable to take them in the form of daily drinks or supplements for preventing or improving obesity. Therefore, there is a need for an energy metabolism-promoting composition containing a component effective in activating PPARγ that can be safely taken daily without relying on such drugs, and a method for activating adipose tissue using the same.

[0015] In addition to the above effects, when ingested as an energy metabolism-promoting composition, the proportion of the active ingredient contained therein that escapes degradation by gastric acid and is released into the bloodstream after undergoing the first-pass effect (removal rate) in the small intestine and liver, i.e., bioavailability, is an important factor. It is known that bioavailability is significantly affected by various ingredients ingested during meals when taking drugs. Insoluble cellulose, for example, has the effect of protecting various drugs from gastric acid, but is also known to reduce their absorption from the small intestine. Conversely, fatty acids such as medium-chain fatty acids are known to promote absorption from the digestive tract. Therefore, for an energy metabolism-promoting composition that has such effects in the form of a beverage or supplement that can be taken daily to prevent or improve obesity, it is important to increase the bioavailability of the active ingredient contained therein and maintain it within an effective range.

[0016] Furthermore, Non-Patent Document 5 discloses that when mice fed a high-fat diet were given yogurt containing trigonelline, fat accumulation in adipose tissue was prevented, and that mice fed yogurt containing trigonelline exhibited suppressed organ weight gain and reduced fat accumulation and hypertrophy in white adipocytes. However, although it is understood that weight reduction was achieved by combining trigonelline with yogurt, i.e., by combining yogurt with trigonelline as a control, there is no disclosure or obvious evidence that trigonelline causes energy metabolism by beigeifying white adipocytes, which store excess energy stored as neutral fat. It is also possible that trigonelline acts as a prebiotic, or that consumption of yogurt changes digestion and absorption efficiency. Furthermore, it is also stated that the organ weights of the mice fed yogurt containing trigonelline (HT group) were lower than those shown in Figures 1B and 1C in Non-Patent Document 5, which is one of the reasons for the relative decrease in organ weight (page 4, right column, lines 7-10 of the same document), and it is unclear whether the increase in organ weight is suppressed in the mice fed yogurt containing trigonelline.

[0017] Based on the above, measures that are thought to be effective in preventing or ameliorating obesity include (1) converting white adipocytes to a beige color, (2) anti-inflammatory effects, (3) PPARγ activation, and (4) improving bioavailability, thereby promoting energy metabolism and preventing or ameliorating obesity. However, specific compositions and methods for achieving these goals are currently needed.

[0018] Japanese Patent No. 6259146 Japanese Patent Application Laid-Open No. 2005-124527 Japanese Patent Application Laid-Open No. 2022-538585 Japanese Patent Application Laid-Open No. 2018-030806

[0019] N. Mohamadi et al., “A review on biosynthesis, analytical techniques, and pharmacological activities of trigonelline as a plant alkaloid.”, Journal of Dietary Supplements, 15(2), pp.207-222 (2018).M. Sasaki et al., “Characteristic analysis of trigonelline contained in Raphanus sativus cv. Sakurajima daikon and results from the first trial examining its vasodilator properties in humans.”, Nutrients, 12(6), 1872 (2020).M. Choi et al., “Trigonelline induces browning in 3T3‐L1 white adipocytes. Phytotherapy Research”, 35(2), pp.1113-1124 (2021).N. Konstantinidis et al., “Risk Assessment of Trigonelline in Coffee and Coffee By-Products.”, Molecules, 28(8), 3460 (2023).Mariana Campos Costa et al.,” Trigonelline and curcumin alone, but not in combination, counteract oxidative stress and inflammation and increase glycation product detoxification in the liver and kidney of mice with high-fat diet-induced obesity”, J Nutr Biochem, Feb (2020).

[0020] In light of the above circumstances, the present invention aims to provide an energy metabolism-promoting composition and an energy metabolism promoter that are effective in improving and preventing obesity, and a method for activating adipose tissue using them. In particular, regarding the physiological functions beneficial to health brought about by coffee, many reports have been published on the effects of caffeine, chlorogenic acids, etc., but there have been few reports on trigonelline, and there has been almost no research on its effectiveness on energy metabolism and lipid metabolism in humans. Therefore, the present invention aims to provide an energy metabolism-promoting composition and an energy metabolism promoter that are effective in improving and preventing obesity through the trigonelline contained in coffee, and a method for activating adipose tissue using coffee and an energy metabolism promoter.

[0021] To solve the above problems, the energy metabolism-promoting composition of the present invention contains trigonelline as an active ingredient and has the function of promoting energy metabolism through trigonelline. In particular, as a mechanism of action, trigonelline promotes energy metabolism by promoting the beige coloration of white adipose tissue. The inclusion of trigonelline provides the effects of improving obesity and preventing obesity. The energy metabolism-promoting composition of the present invention may also contain trigonelline derived from coffee. The energy metabolism-promoting composition of the present invention may further contain caffeine to enhance the expression of UCP-1. The energy metabolism-promoting composition of the present invention may also contain chlorogenic acids to exert an anti-inflammatory effect. The fact that chlorogenic acids themselves have an anti-inflammatory effect is also described, for example, in "Muhammad Naveed et al., "Chlorogenic acid (CGA): A pharmacological review and call for further research," Biomedicine & Pharmacotherapy, Volume 97, January 2018, Pages 67-74." The energy metabolism-promoting composition of the present invention may also contain a lipid component to promote gastrointestinal absorption of trigonelline. In the energy metabolism-promoting composition of the present invention, trigonelline is preferably extracted from green coffee beans, but may also be extracted from roasted coffee beans. Extraction from green coffee beans results in a greater amount of trigonelline than extraction from roasted coffee beans. However, extraction from roasted coffee beans has higher extraction efficiency. Trigonelline is preferably contained in an extract of green coffee beans at a concentration of 5 to 25% by mass, and a trigonelline-rich green coffee bean extract is preferable. More preferably, trigonelline is contained in an extract of green coffee beans at a concentration of 8 to 23% by mass. Furthermore, in the energy metabolism-promoting composition of the present invention, trigonelline extracted from green coffee beans may be a green coffee bean extract from which chlorogenic acids and caffeine have been substantially removed.

[0022] The energy metabolism enhancer of the present invention comprises any one of the above-described energy metabolism enhancer compositions. The energy metabolism enhancer of the present invention may be in the form of a beverage, food, supplement, or pharmaceutical product, thereby improving user convenience.

[0023] The brown adipose tissue activator of the present invention comprises any one of the above-mentioned energy metabolism-promoting compositions.

[0024] The agent for promoting beige coloration of white adipose tissue of the present invention comprises any one of the above-mentioned energy metabolism-promoting compositions containing coffee-derived trigonelline.

[0025] The method for activating adipose tissue of the present invention uses any one of the energy metabolism-promoting compositions containing coffee-derived trigonelline to promote beige coloration of white adipose tissue and promote energy metabolism. In the method for activating adipose tissue of the present invention, trigonelline is ingested at 100 to 200 mg / day in humans. Here, in the method for activating adipose tissue of the present invention, the coffee bean extract may be an extract from which chlorogenic acids and caffeine have been further removed. In the method for activating adipose tissue of the present invention, the human has a BMI of 23 kg / m ２ More than 30kg / m ２ The target population is adults under 18 years of age. It has been confirmed that it can improve resting energy expenditure.

[0026] The energy metabolism-promoting composition, the energy metabolism-promoting agent, and the method for activating adipose tissue of the present invention have the effect of improving obesity or preventing obesity.

[0027] Weight transition graph from animal testing Organ weight measurement results from animal testing (1) Organ weight measurement results from animal testing (2) Breath measurement results from animal testing Genetic analysis results from animal testing White fat droplet size measurement results from animal testing Image of fat droplets in subcutaneous white fat in the groin Image of fat droplets in white fat around the testicles Protein analysis results from animal testing Breath measurement results from human testing Brown adipose tissue density measurement results from human testing BMI was 23 kg / m ２ More than 30kg / m２ Actual measured values ​​of resting energy expenditure in subjects with a BMI of less than 23 kg / m ２ More than 30kg / m ２ Changes in brown fat density in subjects under 18 years of age

[0028] The energy metabolism-promoting composition of the present invention containing trigonelline as an active ingredient will be described below. In particular, coffee-derived trigonelline will be described. Coffee-derived trigonelline refers to a composition containing trigonelline as the main component, which is obtained by extraction from components contained in the fruit (coffee cherries) harvested from coffee trees and / or the leaves and flowers of coffee trees. Coffee trees are plants belonging to the genus Coffea in the family Rubiaceae, with Coffea arabica and Coffea robusta being particularly preferred. The various components in the coffee cherries harvested from coffee trees, as well as the leaves and flowers, contain various components in addition to trigonelline. The coffee-derived trigonelline that can be used in the present invention is characterized by containing various proportions of other components in addition to trigonelline.

[0029] Coffee cherries contain parchment (endocarp), silverskin, and mucilage between the flesh covered by an outer skin and the green bean (seed). Green coffee beans typically refer to the dried seeds alone, but in the present invention, trigonelline contained in components other than the seeds can also be used. Furthermore, trigonelline contained in the leaves and flowers of coffee trees can also be used. The components of coffee cherries harvested from these various coffee trees and the amount of trigonelline contained in the leaves and flowers are described in detail in Reference 1: N. Konstantinidis et al., "Risk Assessment of Trigonelline in Coffee and Coffee By-Products," Molecules, 28(8), 3460 (2023). Furthermore, the amount of trigonelline contained in various types of coffee seeds is described in detail in Reference 2: P. Mazzafera, "Trigonelline in coffee. Phytochemistry," 30(7), 2309-2310 (1991). As described in the aforementioned non-patent document, coffee beans contain a maximum of about 7.2 g / kg of trigonelline, while other components other than coffee beans contain a maximum of 62.6 g / kg of trigonelline. Therefore, it is possible to preferably obtain the coffee-derived trigonelline of the present invention from these components as well.

[0030] As the coffee-derived trigonelline of the present invention, trigonelline obtained by extracting roasted beans obtained by roasting green coffee beans can also be preferably used. The amount of trigonelline contained in various roasted coffee beans is described in Reference 3, "H. Taguchi et al., "Trigonelline content in coffee beans and the thermal conversion of trigonelline into nicotinic acid during the roasting of coffee beans," Agricultural and Biological Chemistry, 49(12), pp. 3467-3471 (1985)." Although the amount of trigonelline decreases due to its conversion to nicotinic acid and other substances during roasting, the coffee-derived trigonelline of the present invention can be preferably added to a coffee beverage obtained by extraction from roasted beans, thereby forming an energy metabolism-promoting composition containing the coffee-derived trigonelline of the present invention.

[0031] The energy metabolism-promoting composition of the present invention contains coffee-derived trigonelline as an active ingredient. The coffee-derived trigonelline is characterized by the inclusion of various coffee-derived components in addition to trigonelline. Among these, caffeine is preferred. Coffee beans contain approximately 1-3% caffeine. Caffeine has a sympathomimetic effect, and has been expected to promote fat burning similar to capsaicin. However, recent research, such as Reference 4 (K. Velickovic et al., "Caffeine exposure induces browning features in adipose tissue in vitro and in vivo," Scientific Reports, 9(1), pp. 1-11 (2019)), has shown that the addition of caffeine to white adipocytes induced to differentiate from stem cells increases UCP-1 expression in association with beige coloration. Furthermore, coffee's effects on humans, such as an increase in supraclavicular body temperature associated with the activation of brown adipose tissue, have been observed.

[0032] Therefore, the energy metabolism-promoting composition of the present invention comprising coffee-derived trigonelline can preferably contain caffeine. However, because caffeine has a central nervous system stimulating effect, excessive intake of caffeine or elderly people may cause symptoms such as increased heart rate and blood pressure, as well as insomnia due to associated excitement and anxiety. In such cases, it is preferable to use the caffeine-free energy metabolism-promoting composition of the present invention.

[0033] The energy metabolism-promoting composition of the present invention can be preferably used when it contains chlorogenic acids as components contained together with coffee-derived trigonelline. Chlorogenic acids are polyphenols containing 5-caffeoylquinic acid, and are contained in coffee beans at approximately 5-11%. Chlorogenic acids have been approved as active ingredients in functional foods, where their efficacy in reducing body fat and blood pressure has been recognized. While the ingestion of chlorogenic acids has been recognized as a fat-burning promoting effect, its mechanism of action differs from that of trigonelline. It is believed that continuous ingestion of chlorogenic acids increases the transport of fatty acids taken up into the liver into mitochondria, thereby promoting fatty acid combustion within the liver.

[0034] On the other hand, it is known that some chlorogenic acids are converted to caffeic acid in the small intestine and then appear in the blood as sulfate conjugates of caffeic acid and ferulic acid. Furthermore, after being converted to dihydrocaffeic acid in the large intestine, they appear in the blood as sulfate conjugates. Chlorogenic acids do not appear in the blood in their original structure, but are metabolized and transported to various tissues in the body as carboxylic acid-containing phenolic derivatives, such as caffeic acid, ferulic acid, and dihydrocaffeic acid, for absorption. In the present invention, we focused on the excellent antioxidant properties of various phenolic compounds derived from these chlorogenic acids and found that imparting an anti-inflammatory effect, particularly in white adipose tissue, that suppresses inflammation induced by macrophage infiltration due to obesity, to the energy metabolism-promoting composition of the present invention is effective in preventing and ameliorating obesity. Therefore, chlorogenic acids can be preferably included in the energy metabolism-promoting composition of the present invention, which comprises coffee-derived trigonelline. However, since chlorogenic acids can bind to metal ions such as zinc and inhibit the absorption of zinc from the digestive tract, in cases where symptoms such as zinc deficiency are present, it is preferable to use the energy metabolism-promoting composition of the present invention, which does not contain chlorogenic acids.

[0035] The energy metabolism-promoting composition of the present invention can be preferably used when it contains lipid components as components contained in addition to coffee-derived trigonelline. It is known that both green coffee beans and roasted coffee beans contain approximately 10 to 20% lipid components. The compounds contained as lipid components include large amounts of triglycerides such as linoleic acid, palmitic acid, stearic acid, and oleic acid, but in addition, diterpenes such as cafestol and kahweol and their ester compounds account for approximately 20% of the total lipids. Furthermore, the surface of green coffee beans contains a lipid component known as coffee wax, which contains tryptamin esters formed by esterifying serotonin with fatty acids.

[0036] These various coffee-derived lipid components are expected to promote the gastrointestinal absorption of the above-mentioned coffee-derived trigonelline and the chlorogenic acids contained therein, thereby improving the bioavailability of these active ingredients. Because these lipid components are insoluble in water, they can be extracted from coffee by adding an alcohol in addition to water, and then used together with coffee-derived trigonelline in the energy metabolism-promoting composition of the present invention.

[0037] The energy metabolism-promoting composition of the present invention can preferably use the above-mentioned coffee-derived trigonelline extracted from various components of the coffee plant, either alone or in combination with it, in addition to caffeine, chlorogenic acids, and lipid components. However, it is preferable to combine these various components and form them into a formulation such as a solution, powder, or capsule, and use this as an energy metabolism promoter.

[0038] The energy metabolism enhancer preferably contains various other components in addition to the above-mentioned components as constituents thereof. Specifically, for example, p-synephylline, which is a naturally occurring component that selectively acts on the β3-adrenergic receptor (β3AR) and is effective in burning lipids, and is obtained by extraction from mandarin orange peel or the like, may be contained in the energy metabolism enhancer together with the composition.

[0039] Alternatively, ferulic acid, which is contained in rice, wheat, and other crops in addition to coffee beans, is transferred into the body and exhibits anti-inflammatory effects due to its antioxidant properties in various tissues, and is therefore expected to have the effect of suppressing inflammation caused by macrophages infiltrating into white adipose tissue. Therefore, it is also preferably included in the energy metabolism promoter of the present invention.

[0040] Furthermore, many naturally occurring compounds are known to have the effect of activating the aforementioned PPARγ and promoting fat metabolism. For example, gingerol contained in ginger not only stimulates the sympathetic nerve and promotes fat burning like capsaicin, but also activates PPARγ, and is therefore preferably included in the energy metabolism promoter of the present invention. Alternatively, pinellic acid contained in onions is also expected to have the effect of activating PPARγ and promoting fat metabolism, and therefore an onion extract containing this can also be preferably included in the energy metabolism promoter of the present invention. Furthermore, since DHA (docosahexanoic acid) contained in fish oil and the like is expected to activate PPARγ and promote fat metabolism (Reference 5: J. Song et al., "DHA increases adiponectin expression more effectively than EPA at relatively low concentrations by regulating PPARγ and its phosphorylation at Ser273 in 3T3-L1 adipocytes," Nutrition & Metabolism, 14(1), pp. 1-11 (2017)) and thus can be preferably contained in the energy metabolism promoter of the present invention. Furthermore, since propolis extract is known to contain components that activate PPARγ and promote fat metabolism (Reference 6: Miho Aga et al., "Propolis Extract Ameliorates Insulin Resistance in 3T3-L1 Preadipocytes," Journal of the Japanese Society for Food Science and Technology, 56(1), pp. 31-39 (2009)), it can be preferably contained in the energy metabolism promoter of the present invention.

[0041] The use of the above-mentioned various naturally occurring ingredients as the energy metabolism promoter of the present invention, either alone or together with coffee-derived trigonelline, in an energy metabolism-promoting composition containing coffee-derived trigonelline, is expected to promote fat burning and be effective in preventing and improving obesity.

[0042] As a method for activating adipose tissue using the above-mentioned coffee-derived trigonelline alone, an energy metabolism-promoting composition containing coffee-derived trigonelline, or an energy metabolism-promoting agent containing coffee-derived trigonelline, using these together with a coffee beverage is expected to activate adipose tissue and be effective in preventing and improving obesity.

[0043] An example of an embodiment of the present invention will be described in detail below with reference to the drawings. Note that the scope of the present invention is not limited to the following examples and illustrated examples, and many modifications and variations are possible.

[0044] (Animal Testing) Eight-week-old male diet-induced obesity model mice (C57BL / 6J) were orally administered a trigonelline-rich green coffee bean extract (chlorogenic acids, caffeine removed) along with a high-fat diet for 16 weeks (trigonelline 300 mg / kg / day), and body weight, organ weight, resting energy expenditure, genetic analysis, protein analysis, and white fat lipid droplet size were measured. The number of animals used in the test was n=8 per group. The trigonelline content in the extract was 29.2% by mass, and chlorogenic acids and caffeine were below the lower limit of detection (N.D.: Not Detected). Note that the trigonelline shown here is the percentage in terms of trigonelline hydrochloride equivalent, which is 23.0% by mass in terms of molecular weight of trigonelline. Other components include lipids, fiber, etc. The test results are explained with reference to Figures 1 to 9. In each case, Comparative Example A was administered a normal diet, Comparative Example B was administered a high-fat diet, and Example C was administered a high-fat diet and trigonelline at the same time. The specific composition of the sample in Example C is as shown in Table 1 below.

[0045]

[0046] Here, trigonelline, chlorogenic acids, and caffeine in the green coffee bean extract were quantitatively analyzed using HPLC-PDA (photodiode array detector). The analytical method is shown in Table 2 below.

[0047]

[0048] Here, chlorogenic acids is a collective term for nine chlorogenic acids: three CQAs (3-CQA, 5-CQA, 4-CQA), three FQAs (3-FQA, 5-FQA, 4-FQA), and three di-CQAs (3,5-di-CQA, 3,4-di-CQA, 4,5-di-CQA). Trigonelline shown in Table 1 is the equivalent of trigonelline hydrochloride used as a standard sample.

[0049] (Regarding body weight) Figure 1 shows a graph of body weight transition in the animal test. As shown in Figure 1, the body weight increased in Comparative Example B, which was given a high-fat diet, compared to Comparative Example A, which was given a normal diet, but in Example C, which was given a high-fat diet and trigonelline simultaneously, weight gain was suppressed from about 60 days onwards.

[0050] (Regarding organ weight) Figures 2 and 3 show the results of organ weight measurement in animal tests, with Figure 2 (1) showing liver weight, Figure 2 (2) showing perirenal white fat weight, Figure 3 (1) showing peritestrial white fat weight, and Figure 3 (2) showing inguinal subcutaneous white fat weight. As shown in Figures 2 and 3, in Comparative Example B, in which a high-fat diet was ingested, liver weight, perirenal white fat weight, peritestrial white fat weight, and inguinal subcutaneous white fat weight all increased compared to Comparative Example A, in which a normal diet was ingested, but in Example C, in which a high-fat diet and trigonelline were ingested simultaneously, weight gain in all of these was suppressed. These findings suggest that simultaneous ingestion of trigonelline has an obesity-suppressing effect.

[0051] (Regarding energy expenditure) In animal tests, resting energy expenditure was calculated by measuring breath. Here, resting energy expenditure is based on the minimum amount of energy required to maintain life, such as maintaining heart rate, breathing, and body temperature, and is known to account for 50-70% of total energy expenditure. Increasing resting energy expenditure, which is the basis for total daily energy expenditure, can improve obesity in animals. Specifically, oxygen consumption (VO2) for approximately 22 hours (12:00 to 10:00 the next day) was measured using an animal energy metabolism measurement system (ARCO-2000N, manufactured by Arco Systems). ２ ) and carbon dioxide emissions (VCO ２ ) was measured. At the same time, spontaneous movement activity was measured using a spontaneous movement activity sensor (NS-AS01, manufactured by Neuroscience). From these results, the respiratory quotient, calorie expenditure (kcal / min), and total energy expenditure were calculated using the following formulas 1 to 4.

[0052] (Equation 1) Respiratory quotient (RQ) = carbon dioxide output (VCO ２ ) / oxygen consumption (VO ２ ) ... (Formula 1)

[0053] (Equation 2) Calories burned = CV x VO ２ ...(Equation 2) CV value (heat generation amount) = 3.815 + 1.232 × RQ ...(Equation 3)

[0054] (Equation 3) Total energy expenditure (kcal / day) = Calorie expenditure × Time (Equation 4)

[0055] FIG. 4 shows the breath measurement results for the animal test. The vertical axis represents energy consumption (kcal / day) per 100 g of body weight, and the measurement results for Comparative Example A, Comparative Example B, and Example C at 4, 6, 12, and 16 weeks after the start of the test are shown. As shown in FIG. 4, Comparative Example B, which consumed a high-fat diet, showed a greater decrease in energy consumption over time than Comparative Example A, which consumed a normal diet. This indicates that energy consumption decreases as obesity progresses. In contrast, Example C, in which a high-fat diet and trigonelline were consumed simultaneously, showed a more gradual decrease in resting energy consumption than Comparative Example B, which consumed a high-fat diet, indicating an improvement. From the above, it was found that simultaneous intake of trigonelline can suppress the decrease in energy metabolism caused by obesity.

[0056] (Regarding genetic analysis) Cell samples were collected from inguinal white adipose tissue, and total RNA was extracted using the RNeasy Lipid Tissue Mini Kit (QIAGEN). After adjusting the RNA concentration, cDNA was synthesized using the QuantiTect Reverse Transcription Kit. The relative expression level of each gene relative to GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was then quantified by real-time PCR. For each genetic analysis, the three marker genes (mRNA expression levels) of UCP-1, PGC-1α, and PRDM16 were measured. Upregulation of these three marker genes suggests that white fat has become beige fat. Figure 5 shows the results of genetic analysis in animal testing, with (1) showing the relative level of UCP-1 mRNA, (2) showing the relative level of PGC-1α mRNA, and (3) showing the relative level of PRDM16 mRNA. As shown in Figures 5(1) to 5(3), in the genetic analysis of inguinal white fat, Example C, in which a high-fat diet and trigonelline were simultaneously ingested, showed upregulation of all of UCP-1, PGC-1α, and PRDM16 compared to Comparative Example B, in which a high-fat diet was ingested.

[0057] Generally, white fat has a large lipid droplet size, while beige fat has a smaller lipid droplet size than white fat. Therefore, a decrease in lipid droplet size suggests that white adipocytes have been converted to beige adipocytes. Figure 6 shows the results of measuring the lipid droplet size of white fat in an animal test, where (1) shows the lipid droplet size of subcutaneous white fat in the inguinal region, and (2) shows the lipid droplet size of peritesticular white fat. Figure 7 shows an image of lipid droplets in subcutaneous white fat in the inguinal region, and Figure 8 shows an image of lipid droplets in peritesticular white fat, where (1) shows Comparative Example A, (2) shows Comparative Example B, and (3) shows Example C. As shown in Figures 6(1) and (2), in Example C, where a high-fat diet and trigonelline were simultaneously ingested, the lipid droplet size of the inguinal white fat or peritesticular white fat was reduced compared to Comparative Example B, where a high-fat diet was ingested.

[0058] Regarding the image of fat droplets in inguinal white fat, the fat droplet size of Comparative Example B shown in Figure 7(2) is larger than that of Comparative Example A shown in Figure 7(1), whereas the fat droplet size of Example C shown in Figure 7(3) is smaller than that of Comparative Example B. Similarly, regarding the image of fat droplets in peritestril white fat, the fat droplet size of Comparative Example B shown in Figure 8(2) is larger than that of Comparative Example A shown in Figure 8(1), whereas the fat droplet size of Example C shown in Figure 8(3) is smaller than that of Comparative Example B. These findings suggest that simultaneous administration of trigonelline causes beige coloration of white adipocytes.

[0059] (Protein Analysis) For protein analysis of dorsal brown adipose tissue cells, UCP-1 was measured by Western blotting. Proteins were eluted from frozen dorsal brown adipose tissue cells to obtain a tissue lysate, and total protein was quantified using a microplate reader (wavelength 562 nm) together with a solution using the BSA Standard included in the BCA Protein Assay Kit (manufactured by Thermo Fisher Scientific). The tissue lysate was subjected to electrophoresis using an XCell SureLock Mini-Cell (manufactured by Thermo Fisher Scientific), and blotting was performed on a PVDF membrane using a semi-dry type flatbed transfer device. The PVDF membrane was developed on Hyperfilm ECL (manufactured by Cytiva), and the image data was digitized using a CS analyzer (manufactured by Atto Corporation) to determine the amount of target protein. UCP-1 is upregulated by thermogenesis. Figure 9 shows the protein analysis results for animal testing, with the vertical axis showing the relative UCP-1 value. As shown in Figure 9, in the protein analysis of dorsal brown adipocytes, UCP-1 was upregulated in Example C, in which a high-fat diet and trigonelline were simultaneously ingested, more than in Comparative Example B, in which a high-fat diet was ingested. These findings suggest that simultaneous ingestion of trigonelline activates brown adipocytes.

[0060] (Human Testing) Healthy men and women were given a trigonelline-rich coffee bean extract (chlorogenic acids, caffeine removed) orally for 8 weeks (150 mg trigonelline / day), and their resting energy expenditure and brown adipose tissue density were measured. The coffee bean extract material contained 10.6% by mass of trigonelline. Specifically, resting energy expenditure was measured using a pulmonary exercise monitoring system (Aeromonitor AE-310S, manufactured by Minato Medical Science Co., Ltd.) at room temperature of 19°C, based on oxygen consumption (VO2). ２ ) and carbon dioxide emissions (VCO ２ From these results, resting energy expenditure was calculated using the following formula:

[0061] (Equation 4) Resting energy expenditure (kcal / day) = 5.616 × VO ２ (ml / min) + 1.584 × VCO ２ (ml / min) (Formula 5)

[0062] Brown adipose tissue density was measured using a time-resolved spectroscopy system (TRS-21, manufactured by Hamamatsu Photonics) to measure the total hemoglobin content in the supraclavicular fossa at room temperature of 25°C. The test results will be explained with reference to Figures 10 and 11. In both cases, Comparative Example D shows a placebo (fake drug) and Example E shows a case where trigonelline was administered.

[0063] The specific composition of the sample of Example E is as shown in Table 3 below. The analytical values ​​shown in Table 3 were also determined according to the analytical method shown in Table 2. Note that the trigonelline shown in Table 3 is the trigonelline equivalent calculated from the molecular weight of trigonelline hydrochloride used as a standard sample.

[0064]

[0065] (Regarding resting energy expenditure) Figure 10 shows the results of breath measurements in a human study, where (1) shows the resting energy expenditure at weeks 0 and 8, and (2) shows the change in resting energy expenditure at weeks 0 and 8. As shown in Figures 10(1) and (2), in Comparative Example D, where a placebo was administered, almost no change was observed between weeks 0 and 8, but in Example E, where trigonelline was administered, resting energy expenditure increased between weeks 0 and 8.

[0066] (Regarding Brown Adipose Tissue Density) Figure 11 shows the results of brown adipose tissue density measurements in human trials, specifically the results of measurements of supraclavicular brown adipose tissue density. Figure 11 (1) shows the total hemoglobin amount, and (2) shows the change in total hemoglobin amount. As shown in Figures 11 (1) and (2), Example E, which took trigonelline, showed an increase in total hemoglobin amount at 0 and 8 weeks compared to Comparative Example D, which took a placebo, indicating an increase in supraclavicular brown adipose tissue density. From the above, it was suggested that the activation of brown or beige adipocytes improves energy metabolism in human trials.

[0067] (BMI is 23kg / m ２ More than 30kg / m ２ (For human studies in subjects under 20 years of age) Trigonelline-rich coffee bean extract (chlorogenic acids, decaffeinated) was administered to subjects with a BMI of 23 kg / m ２ More than 30kg / m ２ Healthy men and women (nine in the test food intake group and 11 in the placebo intake group) under 8 years of age were given trigonelline (150 mg / day) orally for 8 weeks, and their resting energy expenditure and brown adipose tissue density were measured. The coffee bean extract material contained 8.2% by mass of trigonelline. Specifically, resting energy expenditure was measured using a pulmonary exercise monitoring system (Aeromonitor AE-310S, manufactured by Minato Medical Science Co., Ltd.) to measure oxygen consumption (VO2) at room temperature of 27°C and 19°C. ２ ) and carbon dioxide emissions (VCO ２ From these results, the resting energy expenditure was calculated using the above formula 5.

[0068] Brown adipose tissue density was measured using a time-resolved spectroscopy system (TRS-21, manufactured by Hamamatsu Photonics) to measure the total hemoglobin content in the supraclavicular fossa at room temperature of 27°C. The test results will be explained with reference to Fig. 12 or Fig. 13. In both cases, Comparative Example G shows a placebo (fake drug) and Example F shows a case in which trigonelline was ingested.

[0069] The specific composition of the sample of Example F is as shown in Table 4 below. The analytical values ​​of caffeine, coffee-derived chlorogenic acids, and coffee-derived trigonelline shown in Table 4 were also determined according to the analytical methods shown in Table 2 above. Note that the trigonelline shown in Table 4 is the trigonelline equivalent converted from the molecular weight of trigonelline hydrochloride used as a standard, as in Table 3 above.

[0070]

[0071] (Regarding resting energy expenditure) BMI is 23 kg / m ２ More than 30kg / m ２The results of resting energy expenditure for subjects under 80 years of age (9 subjects in the test food intake group and 11 subjects in the placebo food intake group) are shown in Table 5 below and Figure 12. Table 5 shows values ​​expressed as mean ± standard deviation, with SCR indicating the time point at screening (before intake began). Figure 12(1) shows the change in actual measured values ​​of resting energy expenditure at 27°C over time, and Figure 12(2) shows the change in actual measured values ​​of resting energy expenditure at 19°C over time, expressed as mean ± standard deviation. Note that "*" in both Table 5 and Figure 12 indicates "p<0.05 vs. placebo," and while the values ​​at 27°C and 8W were calculated using partial interpolation using LOCF (Last Observation Carried Forward), neither of these numerical interpolations favored the test food intake group. Significant differences were confirmed between the groups in the actual measured values ​​of resting energy expenditure at 19°C at 4 weeks and at 27°C at 8 weeks, with the test food intake group (Example F) showing higher values ​​compared to the placebo food intake group (Comparative Example G) (4 weeks (19°C): 1779.88±202.84 (kcal / day) vs. 1574.68±315.52 (kcal / day), p=0.044; 8 weeks (27°C): 1841.40±225.45 (kcal / day) vs. 1627.79±254.55 (kcal / day), p=0.044).

[0072]

[0073] (Regarding brown adipose tissue density) BMI is 23 kg / m ２ More than 30kg / m ２ The results of brown adipose tissue density (total hemoglobin amount) in subjects under 8 years of age (9 subjects in the test food intake group, 11 subjects in the placebo food intake group) are shown in Table 6 below and Figure 13. Table 6 shows values ​​as mean ± standard deviation, and SCR indicates the time point at screening (before intake began). Figure 13 illustrates the time course of change in brown fat density as mean ± standard deviation. Note that "#" indicates "p<0.10 vs. placebo" in both Table 6 and Figure 13. The change in brown adipose tissue density at 8 weeks showed a significant trend between the groups, with the test food intake group (Example F) showing a higher value (12.31 ± 31.65 (μM) vs. -6.68 ± 22.14 (μM), p = 0.063) compared to the placebo food intake group (Comparative Example G).

[0074]

[0075] In this study, brown fat density was measured in the supraclavicular fossa, and it has been reported that beige adipocytes are present in this area in addition to classical BAT. Therefore, the higher brown adipose tissue density change over eight weeks in the test food intake group suggests that this is due to an increase in beige adipocytes caused by the coffee-derived trigonelline promoting the beige coloration of white adipose tissue.

[0076] This study was a placebo-controlled, randomized, double-blind, parallel-group comparative study. Subgroup analysis showed that patients with a BMI of 23 kg / m ２ More than 30kg / m ２ The resting energy expenditure and brown adipose tissue density of subjects with a BMI of 23 kg / m were analyzed, and the randomization was not disrupted. ２ More than 30kg / m ２ It was shown that adipose tissue activation promoted beige transformation of white adipose tissue in subjects under 18 years of age and promoted energy metabolism.

[0077] The present invention is useful as a technique for improving obesity conditions and preventing obesity.

Claims

1. An energy metabolism-promoting composition which contains trigonelline as an active ingredient and promotes energy metabolism through trigonelline.

2. The energy metabolism-promoting composition according to claim 1, which promotes the beigeification of white adipose tissue by trigonelline and promotes energy metabolism.

3. The energy metabolism-promoting composition according to claim 2, wherein trigonelline is derived from coffee.

4. The energy metabolism-promoting composition according to claim 3, further comprising caffeine and enhancing the expression of UCP-1.

5. The energy metabolism-promoting composition according to claim 4, further comprising chlorogenic acids to exert an anti-inflammatory effect.

6. The energy metabolism-promoting composition according to claim 5, further comprising a lipid component to promote gastrointestinal absorption of trigonelline.

7. The energy metabolism-promoting composition according to claim 3, wherein trigonelline is extracted from green coffee beans, and the green coffee bean extract contains 5 to 25% by mass of trigonelline.

8. The energy metabolism-promoting composition according to claim 7, wherein trigonelline is obtained by substantially removing chlorogenic acids and caffeine from a green coffee bean extract.

9. An energy metabolism promoter comprising the energy metabolism promoting composition according to any one of claims 1 to 8.

10. The energy metabolism promoter according to claim 9, which is in the form of a beverage, food, supplement or medicine.

11. A brown adipose tissue activator comprising the energy metabolism promoting composition according to any one of claims 1 to 8.

12. An agent for promoting beige coloring of white adipose tissue, comprising the energy metabolism-promoting composition according to any one of claims 3 to 8.

13. A method for activating adipose tissue, which comprises promoting the beigeification of white adipose tissue and promoting energy metabolism, using the energy metabolism-promoting composition according to any one of claims 3 to 8.

14. The method for activating adipose tissue according to claim 13, wherein the trigonelline is ingested by a human at a dose of 100 to 200 mg / day.

15. The human has a BMI of 23 kg / m ２ More than 30kg / m ２ The method for activating adipose tissue according to claim 14, which is intended for adults under 20 years of age.

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