JPWO2005082390A1 - Fat accumulation inhibitor - Google Patents

Abstract

An object of the present invention is to provide a fat accumulation-inhibiting agent which is derived from a natural product and does not adversely affect the human body and has an effect of inhibiting fat accumulation per body weight. The present invention relates to a fat accumulation inhibitor containing, as an active ingredient, a fruit polyphenol derived from a plant, preferably a rose family plant, particularly an apple fruit or an immature fruit. Specifically, the polyphenol is a proanthocyanidin, particularly procyanidin. Yes, foods, beverages, food additives, pharmaceuticals, quasi drugs, and feeds containing them.

Description

The present invention relates to a fat accumulation inhibitor containing, as an active ingredient, a fruit polyphenol derived from a plant, preferably a Rosaceae plant, particularly an apple fruit or an immature fruit. Specifically, the polyphenol is proanthocyanidin, particularly procyanidin, The present invention relates to a fat accumulation inhibitor having effects such as inhibition of insulin elevation, inhibition of leptin elevation, and suppression of fat accumulation per body weight, and foods, beverages, food additives, pharmaceuticals, quasi drugs, and feeds containing the same.

Western meals as well as Japanese meals have been remarkably westernized in recent years, and high-fat and high-calorie diets are progressing. In particular, excessive intake of fat and sugar causes fat accumulation, and an increasing number of people suffer from obesity and increased body fat. Body fat is the sum of subcutaneous fat and visceral fat, and a positive correlation is obtained between visceral fat mass and body fat, and it is known that visceral fat mass reflects body fat.
Obesity and increased body fat tend to be associated with hypertension and hyperlipidemia, and are considered risk factors such as ischemic heart disease, stroke, diabetes, etc. Countermeasures for obesity are extremely important from the viewpoint of lifestyle-related disease prevention It is.
Most obesity prevention and treatment methods used so far have been dietary restrictions and dietary diets, but dietary restrictions are accompanied by mental difficulties that can lead to pathological symptoms such as malnutrition and anorexia. There is also danger. In addition, excessive diet and inhibition of digestive enzymes reduce the supply of nutrients in the body, causing rebound after dieting.
On the other hand, various pharmaceuticals are also administered, but side effects must be taken into account along with their efficacy.

Here, polyphenols widely present in the plant kingdom have been confirmed to have various physiological functions by recent research. Polyphenols derived from rosaceae fruits or immature fruits can also be used for antioxidant, anti-cariogenic, allergy suppression, etc. A function has been found (Patent Document 1), but no function for inhibiting fat accumulation has been reported.
In vivo lipid metabolism suppression using fruit polyphenols as an active ingredient has been reported (Patent Document 2). In vivo lipid oxidation suppression, HDL (good cholesterol) / total cholesterol ratio improvement, cholesterol absorption suppression Although it has been reported, there is no mention of the relationship between fruit polyphenols and procyanidins and fat accumulation.
Although procyanidin has been reported for prevention of obesity (Patent Document 3), the examples show weight loss and suppression of serum lipids and blood sugar elevation due to eating disorders or digestive enzyme inhibition, and the amount of body fat per body weight No mention is made of (visceral fat mass).
JP-A-7-285876 Japanese Patent Laid-Open No. 10-330278 JP-A-9-291039

As described above, a fat accumulation inhibitor that does not affect body weight gain and has an effect of suppressing increase in insulin and leptin in vivo, particularly fat accumulation per body weight, prevents increase in body fat percentage, It is expected to prevent hyperlipidemia, ischemic heart disease, stroke, diabetes and the like.
Therefore, an object of the present invention is to provide a fat accumulation inhibitor satisfying these properties, which is derived from a natural product and does not adversely affect the human body.

As a result of intensive studies to find a natural product that has a fat accumulation inhibitory action and is safe in a high fat diet system, the present inventors have found that polyphenols derived from apple fruits or immature fruits are insulin in vivo. It has been found that it has an inhibitory effect on elevation, an inhibitory effect on elevation of leptin, particularly an inhibitory effect on fat accumulation per body weight, and has reached the present invention.
That is, according to the present invention, there is provided a fat accumulation inhibitor characterized by containing a polyphenol derived from a fruit or an immature fruit as an active ingredient.
In addition, the fat accumulation inhibitor of the present invention suppresses fat accumulation per body weight by suppressing increase in insulin and leptin in vivo.
The fat accumulation inhibitor of the present invention is preferably a polyphenol derived from a rose family plant, especially an apple fruit or an immature fruit.
Furthermore, the fat accumulation inhibitor of the present invention can be used in foods, beverages, food additives, pharmaceuticals, quasi drugs and feeds.

According to the present invention, there is provided a fat accumulation inhibitor characterized by containing, as an active ingredient, an apple extract or polyphenol or procyanidin extracted / purified from a rose family plant, particularly an apple fruit or an immature fruit, It is possible to suppress fat accumulation by virtue of functions such as inhibition of visceral fat accumulation, hyperinsulinemia, hyperleptinemia and the like possessed by the fat accumulation inhibitor.
Furthermore, the fat accumulation inhibitor of the present invention is a food and beverage, a food additive, a pharmaceutical, and a quasi-drug for those who are concerned about increased body fat, obesity, hyperlipidemia, arteriosclerosis, diabetes and the like. And can be used in animal feed.

It is a graph which shows the mesenteric fat accumulation inhibitory effect by animal experiment using apple polyphenol. It is a graph which shows the total visceral fat accumulation inhibitory effect by animal experiment using apple polyphenol. It is a graph which shows the blood insulin concentration rise inhibitory effect by an animal experiment using apple polyphenol. It is a graph which shows the blood leptin density | concentration raise inhibitory effect effect by animal experiment using an apple polyphenol.

The polyphenol used in the present invention is composed of a polyphenol fraction extracted and purified from a fruit or an immature fruit. The polyphenol fraction is first obtained by squeezing fruit juice of an original fruit or an organic solvent such as water or ethanol. The extracted extract can be used.
The extracted juice from which the juice juice or organic solvent has been removed can be used as it is, but through a process such as centrifugation or filtration, a clear juice or extract is obtained, and the polyphenol fraction further comprises Further, it is purified by an ion exchange resin, a synthetic adsorption resin, an adsorbent such as silica gel, a gel filtration agent, or the like.
The polyphenol fraction can then be concentrated to obtain a liquid formulation, and further, a powder formulation can be obtained by spray drying or freeze-drying the concentrated solution.
An auxiliary agent such as an organic solvent such as ethanol or an organic acid such as citric acid can be used for the liquid preparation, and a dextrin, saccharide or the like can be used as an excipient for the powder preparation.
In the present invention, fruits, preferably fruits of Rosaceae, specifically apples, pears, peaches, umes, etc. can be used as raw materials, but apples are particularly preferred because of their high polyphenol concentration.
Moreover, although a mature fruit and an immature fruit can be used as a fruit, an immature fruit is especially preferable from containing many polyphenols and many active ingredients which have a wide range of physiological functions.
The dose when the fat accumulation inhibitor, the insulin elevation inhibitor, and the leptin elevation inhibitor described in the present invention are used as an oral preparation varies depending on the purpose of administration and the sex, age, weight, health status, etc. of the administration subject. However, the dry polyphenol fraction can be administered in the range of 1 mg / kg body weight to 100 mg / kg body weight.
The polyphenol derived from the apple of the present invention has no problem in terms of safety because the raw material is a fruit used in food, but for the polyphenol fraction, an acute toxicity test, a subchronic toxicity test, a mutagenicity test, etc. Various safety tests are conducted to confirm its safety.
The polyphenol fraction obtained as described above contains caffeic acid derivatives (esters) such as chlorogenic acid, p-coumaric acid derivatives, flavon-3-ols (catechins), flavonols such as quercetin glycosides, and phloretin. It is composed of chalcones such as glycosides, polyphenols such as procyanidins such as procyanidin B-2, among others, a composition containing procyanidins as main components, and antioxidant, suppression of blood pressure increase, suppression of mutagenicity, It has already been confirmed that it has physiological functions such as allergic suppression, anti-caries, deodorization, and in vivo lipid metabolism control (JP-A-7-285876, JP-A-10-330278).

Thereafter, as a result of intensive studies by the present inventors, it has been confirmed that the polyphenol has the following new physiological functions.
First, it was confirmed that the polyphenol has a function of suppressing the accumulation of visceral fat. That is, the polyphenols containing the procyanidins as main components are extremely effective for suppressing body fat accumulation.
Second, it was confirmed that the polyphenol has a function of suppressing an increase in insulin. That is, the polyphenol containing procyanidins as the main component is extremely effective for improving hyperinsulinemia.
Thirdly, it was confirmed that the polyphenol has a function of suppressing an increase in leptin. That is, the polyphenol containing the procyanidins as main components is extremely effective in improving hyperleptinemia.
EXAMPLES Next, although this invention is demonstrated further in detail based on an Example, this invention is not limited to these Examples.

After three weeks old Sprague-Dawley (SD) male rats were preliminarily raised on a commercial solid feed for one week, a high-fat diet based on the diet composition of obesity and diabetes-induced diet (control is a low-fat diet) A feed containing 1.0% of the polyphenol (no blend in the control group) was fed for 64 days. In addition, since the intake of polyphenol-containing feed temporarily decreases in the initial administration due to the inherent bitter taste, it is given under the condition of pair feeding of 6 animals per group so that the intake is finally the same in all groups. (Table 1). After completion of the breeding, blood was collected from the abdominal aorta under ether anesthesia, blood and various tissues were collected, and various fat and tissue weights and various parameters of lipid metabolism system of liver and serum were measured.

Adipose tissue weight: After rearing, the peri-renal (retroperitoneal), testicle-peripheral (cold testicle) and mesenteric adipose tissue were collected and weighed.
Measurement of serum insulin and leptin: Serum insulin and leptin levels as indicators of obesity were measured using a commercially available ELISA kit (Insulin: ELISA kit from Mercodia; Leptin: Yanaihara Institute ELISA kit).
Statistical processing: All results obtained are expressed as mean ± standard error, and the significance test between each group was performed using Duncan's new multiple range test. The significance level was set at p <0.05.
As a result, as shown in Table 2, weight gain and food intake were significantly increased in the high fat diet group and increased in the high fat diet group as compared to the control group. Although no significant difference was observed between the groups, the visceral fat mass was significantly suppressed as shown in Table 3 and FIGS. 1 and 2.
When looking at the fat accumulation rate (total visceral fat mass increase / control group total visceral fat mass), the high fat diet group is (7.36−4.94) ÷ 4.94 × 100 = 49.0% On the other hand, in the high fat diet + polyphenol group, (5.99−4.94) ÷ 4.94 × 100 = 21.2%, the fat accumulation rate was suppressed, and the change in body fat percentage (total visceral fat The amount of increased fat / weight gain × 100) is (7.36−4.94) ÷ 424.7 × 100 = 0.56% in the high fat diet group, whereas high fat diet + polyphenol In the group, the body fat percentage is also reduced at (5.99−4.94) ÷ 432.7 × 100 = 0.24%.

As a result of the measurement, as shown in FIGS. 3 and 4, the serum insulin concentration and the serum leptin concentration were significantly increased in the high fat diet group as compared to the normal diet control group. The tendency for the rise to be suppressed by the addition of polyphenol was observed.

(Production Example) 300 kg of apple fruit from Aomori Prefecture was crushed and pressed to obtain 210 kg of fruit juice. The obtained fruit juice was clarified with 30 ppm of pectinase, and after centrifugation, clarification was performed by filtration through diatomaceous earth (silica 300S, manufactured by Chuo Silica Co., Ltd.) to obtain a clarified fruit juice. The clarified fruit juice was passed through an adsorbing power ram (Seba Heats HP-20, manufactured by Mitsubishi Chemical Corporation) to adsorb polyphenols. Subsequently, pure water was passed through to remove non-adsorbing substances (such as saccharides and organic acids) in the column, followed by elution with 80% alcohol. Alcohol was concentrated under reduced pressure from the obtained fraction to prepare about 2 kg of an extracted powder product. The extracted powder product was assayed using reversed-phase high-performance liquid chromatography. Chlorogenic acids (about 20%), fluorethylene glycosides (about 5%), flavonols (about 15%), proanthocyanidins (about 50%) ) And other browning substances (about 10%). Furthermore, this protosiadinin reliance is based on the results of analysis with a matrix-assisted laser ionization-time-of-flight mass spectrometer (MALDI-TOF / MS, manufactured by Applied Biosystems). It was confirmed to be an oligomer or polymer from the body to the 15-mer. (M. Ohnishi-kameyama et al. Mass Spectrometry, 11, 31-36, 1997)

(Comparative example) Tablet 140 g of lactose and 17 g of corn starch were mixed, and this mixture was granulated with a paste prepared in advance from 70 g of corn starch. 1 g of magnesium stearate was added to the obtained granules and mixed well, and the mixture was tableted with a tableting machine to produce 1000 tablets.
(Practical example) Tablet 150 g of apple polyphenol obtained in Production Example 1, 90 g of lactose and 17 g of corn starch were mixed, and this mixture was granulated with a paste prepared in advance from 70 g of corn starch. 1 g of magnesium stearate was added to the obtained granules and mixed well, and the mixture was tableted with a tableting machine to produce 1000 tablets.
(Test example) Inhibition of fat accumulation by apple polyphenol (human)
Next, a clinical trial examined the action of apple polyphenols to suppress fat accumulation.

(Method)
Among the subject paid volunteers, 50 males and females over 20 years old who volunteered to participate in this study were selected from 50 men with a body mass index of 22 <BMI ≦ 30 in a preliminary study conducted 4 weeks before the start of the study. Selected. However, those who are taking medicines and health foods that may affect lipid metabolism, those who have a history of food allergies, 200 ml within the first month of this study, or 400 ml within three months Exceeded blood collection (those who donated blood, etc., those who were judged unsuitable for study participation by the study investigator were excluded from the subject. Blood, physical examination) and CT results obtained before ingestion, age, weight, height, body mass index, blood pressure, neutral fat, total fat area (TFA), visceral fat area (VFA) and The study was divided into two groups so that the backgrounds of subcutaneous fat area (SFA), waist / hip, etc. were aligned, and this study was subject to the objectives of the Declaration of Helsinki, and subjects were fully explained about the research content and methods. There was carried out with the consent in writing.

Test meal
The test meal was a tablet containing apple polyphenol produced in a practical example (150 mg / 1 tablet) and a control meal containing no polyphenol produced in a comparative example (placepo tablet). In addition, before conducting the test, the investigator confirmed that there was no distinction between the test beverage and the placebo tablet due to sensory aspects such as flavor and aroma and packaging.
Ingestion method and schedule
The study was a parallel two-group comparative study that adopted a double-blind method with random assignment. The test period was set to a total of 20 weeks, with an observation period of 4 weeks, an intake period of 12 weeks, and an observation period of 4 weeks after the end of intake. Subjects were divided into two groups and ingested test tablets in the following manner.
Group A: Apple polyphenol-containing test tablets (150 mg / tablet) each 4 tablets before dinner (4 tablets / day in total), hereinafter Apple polyphenol test tablet intake group B: Placebo tablets (150 mg / tablet) Before taking 4 tablets each (total 4 tablets / day), the following placebo tablet ingestion group In addition to taking test tablets every day, the subjects had their daily diet, smoking and exercise. Instructed not to change.

Abdominal fat analysis by CT scan was performed on all subjects of CT examination. The analysis was performed using the visceral fat measurement PC software “Fat Scan” (manufactured by N2 System Co., Ltd.) from the image obtained from the abdominal CT tomography of the L4 / L5 intervertebral disc, and the total fat area (TFA), the visceral fat area (VFA) was determined.
The implementation period was a total of two times: the start of intake and 12 weeks after intake (end of intake). In principle, the test was carried out within 4 days before and after the test, and on the test day, the test was performed after at least 4 hours of fasting and 2 hours of fasting.
The results are shown in Table 4. As a result of two-way analysis of variance, the transition of total fat area (TFA), visceral fat area (VFA), and subcutaneous fat area (SFA) in all subjects showed an increase trend in TFA in the placebo tablet ingestion group after 12 weeks of ingestion. (P <0.1), but no variation tendency was observed in the apple polyphenol test tablet intake group.

As for VFA, there was no change tendency in the placebo tablet intake group, whereas a significant decrease tendency was observed 12 weeks after the intake in the apple polyphenol test tablet intake group (p <0.1). As a result of the change in the amount of change, there was a tendency that there was a difference between the placebo tablet intake group and the apple polyphenol test tablet intake group feeling (p <0.1).
From the above results, it was shown that apple polyphenol has an effect of reducing body fat.

Claims (14)

A fat accumulation inhibitor comprising a polyphenol derived from a fruit or an immature fruit as an active ingredient. The fat accumulation inhibitor according to claim 1, wherein the fruit is a Rosaceae plant. The fat accumulation inhibitor according to claim 2, wherein the Rosaceae plant is an apple. The fat accumulation inhibitor according to claim 1, wherein the polyphenol is proanthocyanidin. The fat accumulation inhibitor according to claim 4, wherein the proanthocyanidins are procyanidins. The fat accumulation inhibitor according to any one of claims 1 to 5, which has an inhibitory effect on insulin elevation or leptin elevation in a living body. The fat accumulation inhibitor according to any one of claims 1 to 5, wherein the fat accumulation rate is suppressed. The fat accumulation inhibitor according to any one of claims 1 to 5, which reduces body fat percentage. The foodstuff containing the fat accumulation inhibitor as described in any one of Claims 1-8. A beverage containing the fat accumulation inhibitor according to any one of claims 1 to 8. The food additive containing the fat accumulation inhibitor as described in any one of Claims 1-8. The pharmaceutical containing the fat accumulation inhibitor as described in any one of Claims 1-8. A quasi-drug containing the fat accumulation inhibitor according to any one of claims 1 to 8. The animal feed containing the fat accumulation inhibitor as described in any one of Claims 1-8.