JP2008222656A - Obesity ameliorating and preventing composition and health food - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an obesity ameliorating and preventing composition. <P>SOLUTION: The obesity ameliorating and preventing composition comprises a phenyl glucoside derivative as an active ingredient and a pharmaceutically acceptable excipient. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

The present invention relates to a composition for improving and preventing obesity comprising a phenylglucoside derivative contained in crude sugar as an active ingredient, and a health food.
More specifically, obesity comprising a fraction mainly composed of a phenylglucoside derivative contained in crude sugar and having maximum absorption with respect to ultraviolet rays having a wavelength of 272 nm, and a pharmaceutically acceptable excipient or carrier, particularly The present invention relates to a composition for improving and preventing visceral fat obesity and a health food.

  A method for producing a non-sucrose pigment component extract contained in brown sugar (crude sugar) is obtained by dissolving crude sugar in water and filling the column with activated carbon (Non-patent Document 1) or Amberlite XAD-2, which is a polystyrene resin. It is reported that it is separated and purified by a column chromatography method using a nonpolar adsorbent such as (Patent Documents 1 and 2).

Further, it has been reported that the main component is considered to be 3,4-dimethoxyphenyl-β-D-glucoside or 2,4-dimethoxyphenyl-β-D-glucoside (for example, Non-Patent Document 2). ).
Moreover, it has already been clarified that the above-described pigment component extract has an inhibitory effect on the neutral fat and insulin in serum (for example, Non-Patent Document 2).

  Furthermore, it has also been clarified that the pigment component extract has an antiallergic action, can prevent and improve arteriosclerosis (for example, Patent Document 1), and has effects such as skin activation / whitening action (for example, Patent Document 2).

  On the other hand, in post-war Japan, the number of people with obesity due to the consumption of high-calorie meals due to westernization of diet and lifestyle, overeating, irregular eating habits, and lack of exercise due to diversified transportation methods has increased rapidly. ing.

  Obesity is no longer a matter of style alone. In fact, 80% of people with hyperlipidemia, called lifestyle-related diseases, are obese, 50% of hypertension, 30% of patients with liver disease, and 20% of patients with diabetes are said to be obese. Yes. That is, it is pointed out that obesity is the biggest trigger for lifestyle-related diseases.

As a method for determining the degree of obesity, a physique index (BMI: body mass index) represented by the following equation, which is determined by height and weight, is used.
BMI = weight (kg) / (height (m)) ²
If this BMI is in the following range:
Less than 18.5: Skinny 18.5-25: Appropriate

Less than 25-30: Obesity (once)
30 to less than 35: obesity (twice)
35 to less than 40: obesity (3 degrees)
40 or more: Obesity (4 degrees)
It is judged that.

  However, recently, the body fat percentage, which is the proportion of fat in the body composition, is measured with a body fat meter. If the value is less than 15-20% for men and less than 20-25% for women, it is It is judged as “slightly high” if it is less than 20-25% for men and 25-30% for women, and “high” if it is 25% or more for men and 30% or more for women.

  As another judgment method, if the waist to hip ratio: waist (m) / hip (m) is 0.7 or less in women, it is lower body obesity (pear type), and it should be 0.8 or more. The upper body is obese (apple type), and if it is 1.0 or more in males, it is said to be upper body obesity (apple type). In the case of the above-mentioned pear-type obesity, although it is hardly related to lifestyle-related diseases, apple-type obesity is said to be greatly related to the occurrence of lifestyle-related diseases.

  There are two types of upper body obesity: subcutaneous fat obesity with thick fat around the abdomen and visceral fat obesity with visceral fat firmly, and the latter visceral fat obesity is the majority of the occurrence of lifestyle-related diseases. It is clear that they are deeply involved.

  That is, the habits of lack of exercise accompanying intake of the above-mentioned high-calorie diet, overeating, irregular eating habits, and diversification of transportation means fat accumulation and obesity, insulin resistance diabetes, hyperlipidemia, hypertension It is said to cause so-called lifestyle-related diseases such as arteriosclerosis and metabolic syndrome.

  In recent years, adiponectin has been found as a hormone secreted from adipose tissue. This hormone, when normal, has major effects such as prevention of arteriosclerosis and enhancement of insulin sensitivity, but it has metabolic syndrome and excessive obesity. It is said that when visceral fat accumulates abnormally, the secretion of this adiponectin is suppressed and becomes an important cause of the onset of type 2 diabetes and cardiovascular disease (for example, Non-Patent Document 3).

JP-A-57-88107 Patent 59-48809 Kimura et al., Japanese and Chinese Journal of Medicine, Vol1, No. 1, 1984, 178-179 Kimura et al., Pharmaceutical Journal, 102 (7), 1982, 666-669. Kadowaki T. & Yamauchi T., Endocrine Reviews, 2005, 26, 439

  Many drugs and health foods for improving subcutaneous fat obesity have been found even in the case of the above pear-type obesity and apple-type obesity. However, drugs and health foods that improve and prevent visceral fat, which is the most problematic, are not easy to conduct animal experiments to measure the fat of each organ of the viscera, so there is a scientific support for the built-in fat obesity improvement and prevention drug Currently, few health foods have been developed.

As a result of intensive studies, the present inventors have surprisingly found that a phenylglucoside derivative derived from a crude sugar improves and prevents obesity, particularly visceral fat obesity, and has completed the present invention. .
Thus, according to the present invention, a composition for improving and preventing obesity comprising a phenylglucoside derivative as an active ingredient and a pharmaceutically acceptable excipient is provided.

  According to the present invention, the crude sugar-containing phenylglucoside derivative-containing coloring component that has been conventionally regarded as a waste product in the production process of fine sucrose can be safely used for the improvement and prevention of obesity.

The term “crude sugar” used in the present invention means any of crude sucrose, crude sucrose extract or dried product thereof obtained from sugarcane or sugar radish.
The phenylglucoside derivative used in the present invention is a column chromatography method eluting with water and alcohol described in Patent Documents 1 and 2 or Non-Patent Document 1, or a strongly acidic ion described in Production Example 1 below. Obtained according to any of the column chromatography methods using an exchange resin and eluting with water. However, from the viewpoint that the use of the composition according to the present invention is a drug or health food, the method described in the following Production Example 1 in which no organic solvent is used is most preferable.

  Therefore, the term “phenylglucoside derivative” used in the present invention means any of a fraction containing an eluate containing phenylglucoside derivative separated from crude sugar, a reduced-pressure concentrate of the eluate fraction, and a concentrated dry solid powder.

In addition, the phenylglucoside derivative used for this invention shows the following properties.
(1) The concentrated dry powder is a tan brown slightly hygroscopic powder, soluble in water and alcohol, insoluble in hexane petroleum ether and ether:
(2) A 1% aqueous solution of the concentrated dry powder has a pH of about 7.5:
(3) Infrared absorption spectrum ν _max (Nujol; cm ⁻¹ ): 3300, 1590, 1020 and 720:

(4) Ultraviolet absorption spectrum λ _max (nm) (water): 272, 320:
(5) When 2 to 3 drops of 5% aqueous solution of the above-concentrated dry powder is added to 5 mL boiling boiling reagent solution, a red precipitate is formed. In addition, ferric chloride test solution added to 5% aqueous solution of this product is negative: Furthermore, gelatin test solution added to 5% aqueous solution does not cause precipitation:

(6) Thin layer chromatography:
When 10 mg of the above concentrated dry powder is dissolved in 1 mL of water and tested by the Japanese Pharmacopoeia General Test Method Thin-layer Chromatography under the conditions described below, a single Rf between about 0.6 and 0.85 is used. A marked red spot.
Carrier: Silica gel 60F ₂₅₄ (Merck, thickness 0.25 mm):
Sample attachment amount: 10 μL:
Developing solvent: chloroform: methanol: water (65:35:10) lower layer:
Deployment distance: 10cm:
Detection: After spraying with p-anisaldehyde reagent solution, heating at 105 ° C. for 5 minutes:

(7) Crystallization of 3,4-dimethoxyphenyl-β-D-glucoside from concentrated dry powder:
When the above concentrated dry powder is recrystallized from isopropyl alcohol, a white crystalline powder having a melting point of 154 to 156 ° C. is obtained.
When the crystalline powder was subjected to thin layer chromatography under the conditions described in (6) above, a remarkable red spot appeared in Rf0.8.

Elemental analysis, mass spectrometry and ¹ H-NMR spectrum (400 MHz, CD ₃ OD) of the above crystalline powder were measured and found to be completely consistent with the data of 3,4-dimethoxyphenyl-β-D-glucoside. Powder was confirmed to be 3,4-dimethoxyphenyl-β-D-glucoside.

  In addition, HPLC analysis of the above crystalline powder (column: Phenomenex Synergi 4u Fusion-RP 80A (250 mm × 4.6 mm), mobile phase: 0.1% phosphoric acid aqueous solution; acetonitrile = 95%; 5%, detection wavelength: 282 nm) As a result, it was found that the purity was 96% or more.

  Therefore, the term “phenylglucoside derivative” used in the present invention means either 3,4-dimethoxyphenyl-β-D-glucoside alone or a fraction containing the substance as a main component.

  The inventors of the present invention have included a group in which a high-fat diet was continuously administered to mice twice in the morning and evening for 20 weeks, and a group in which the phenylglucoside derivative was added to a high-fat diet and continuously administered. By measuring the weight difference of liver, subcutaneous adipose tissue, mesenteric adipose tissue, and epididymal adipose tissue and the amount of lipid adhering to the liver, it was found that phenylglucoside derivatives are effective for the improvement and prevention of obesity, especially visceral fat obesity. I found it.

  In addition, blood triglyceride levels, blood total cholesterol levels, and blood free fatty acid levels greatly related to obesity were also measured for each group of mice described above. In each case, phenylglucoside derivatives increased these values. It was clarified to suppress.

  Therefore, according to the present invention, there is provided a composition for improving and preventing visceral obesity comprising a phenylglucoside derivative as an active ingredient, or the derivative and a pharmaceutically acceptable excipient.

  Further, according to the present invention, there is provided a health food comprising a composition for improving and preventing obesity, particularly visceral fat obesity, comprising a phenylglucoside derivative as an active ingredient, or the derivative and a pharmaceutically acceptable excipient. The

  The above-mentioned phenyl glucoside derivative may be used alone, or diluted with a suitable pharmaceutically acceptable excipient, and a powder, granule, tablet, capsule, solution or injection by a method known in the field of manufacturing pharmaceuticals and the like. It can be formulated into various pharmaceutical forms such as an agent.

  Examples of such excipients include binders (e.g. syrup, gum arabic, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone), fillers (e.g. lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine), tablet lubricants ( For example, magnesium stearate, talc, polyethylene glycol or silica), a disintegrating agent (for example, potato starch) or a wetting agent (for example, sodium lauryl sulfate) can be used.

  The tablets may be coated by methods well known in normal pharmaceutical practice. Liquid formulations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, and may be provided as a dry product that is mixed with water or other suitable excipients prior to use. Good.

  Such liquid preparations contain conventional additives such as suspending agents (e.g. sorbitol, syrup, methylcellulose, glucose syrup, gelatin edible fat), emulsifiers (e.g. lecithin, sorbitan monooleate or arabic). Gum), non-aqueous excipients (e.g. almond oil, fractionated coconut oil or glycerin, oily components such as propylene glycol or ethylene glycol), preservatives (e.g. methyl or propyl p-hydroxybenzoate or sorbic acid), and A colorant or a fragrance may be included as desired.

  The above-mentioned health food means a food that is more active than ordinary food, and is intended for health, health maintenance and promotion, for example, liquid or semi-solid, solid product, specifically powder. In addition to granules, tablets, capsules or liquids, confectionery such as cookies, rice crackers, jelly, yokan, yogurt and manju, soft drinks, teas, nutritional drinks, soups and the like can be mentioned.

  In the production process of these foods or to the final product, the composition according to the present invention can be added by mixing, coating, spraying or the like to obtain a health food.

  The composition according to the present invention can be effectively used particularly for humans who fall into the metabolic syndrome, or for humans in the reserve army.

  The daily dose of phenylglucoside derivative to humans for preventing and improving visceral fat obesity and subcutaneous fat obesity is 200 mg to 2000 mg, preferably 300 mg to 1000 mg, and most preferably 300 mg to 600 mg.

  As an administration method, it is desirable to take the composition once or twice a day or after a meal alone or as a composition to which an excipient is added.

  Hereinafter, the composition for improving and preventing visceral obesity according to the present invention will be described in detail with reference to examples. Not what you want.

Production Example 1
1 kg of crude sugar powder was dissolved in 2.5 L of deionized water, and insoluble matter was filtered off to obtain an aqueous solution of crude sugar. This aqueous solution was poured into the upper part of a column packed with 17 L of sodium-type cation exchange resin (UBK530 manufactured by Mitsubishi Chemical) dispersed in deionized water and adsorbed onto the resin.
Next, at room temperature, 85 L of ion exchange water was used to elute sucrose by elution from the above column at a flow rate of 8 L / hour.

  After elution of sucrose, when the eluate from the column started to color, it was fractionated by 4 L to obtain fractions 1-20. Each fraction was measured with a sugar content meter (MASTE, manufactured by Atago Co., Ltd.). Fractions 8 to 16 having a sugar content Prix of 0, no sweetness, and a maximum ultraviolet absorption wavelength of 272 nm and an absorbance of 1.0 or more were combined. The eluate was concentrated under reduced pressure at 60 ° C., and the residue was further concentrated to dryness under vacuum at 60 ° C. and dried to contain a phenylglucoside derivative containing 3,4-dimethoxyphenyl-β-D-glucoside as a main component. 5 g of dried powder was obtained in 0.5% yield.

  By easily recrystallizing this phenylglucoside derivative-containing fraction from isopropyl alcohol, a white crystalline powder of 3,4-dimethoxyphenyl-β-D-glucoside (mp 154-156 ° C) can be easily obtained. Can do.

  The dry powder of a fraction containing a phenylglucoside derivative mainly composed of 3,4-dimethoxyphenyl-β-D-glucoside obtained in Production Example 1 was used as a phenylglucoside derivative in the following Examples.

Example 1
Measurement of visceral fat, subcutaneous fat and blood fat content Visceral fat (liver, small intestine, mesenteric fat), epididymal tissue, spinal subcutaneous fat tissue using mice fed a low or high fat diet The weight measurement of liver and the amount of liver fat and blood fat (blood neutral fat, free fatty acid, total cholesterol) were measured as follows.

Experimental materials and methods (1) Diet composition in low fat diet and high fat diet Diet composition in low fat diet (hereinafter also abbreviated as LL) and high fat diet (hereinafter also abbreviated as HF) given to experimental animals is shown in the table below. Show.

食餌組成は上記の表に示したようにＬＬはバター３％を含有し、ＨＦはバター４５％を含有し、ＨＦにおけるバター含有量は、ＬＬにおけるバターの量の１５倍という高脂肪食となっている。また、ＬＬの総カロリー数は２９２.８kcal/１００gに対し、ＨＦの総カロリーは５３５.８/１００gとなっている。

As shown in the above table, LL contains 3% butter, HF contains 45% butter, and the content of butter in HF is a high-fat diet that is 15 times the amount of butter in LL. ing. Moreover, the total calorie count of LL is 292.8 kcal / 100g, and the total calorie of HF is 535.8 / 100g.

(2) Experimental animals After six weeks old C57BL / 6 mice (purchased from Clea Japan Co., Ltd.) were preliminarily raised for one week, healthy mice were treated with a low fat diet group, a high fat diet group, a high fat diet, and phenylglucoside. It was divided into four groups, a group administered with a derivative (hereinafter abbreviated as PGD) (500 mg / kg) and a group administered with a high fat diet and PGD (2000 mg / kg), and the mice were allowed to eat freely using 7 mice for each group. For 20 weeks.
In addition, the above PGD was dissolved in distilled water at a rate of 500 mg / kg and 2000 mg / kg per day per mouse weight and orally administered to each group of mice.

(3) Effect of phenylglucoside derivative (PGD) administration on the increase in the weight of liver, mesenteric adipose tissue, epididymal adipose tissue and subcutaneous adipose tissue in obese mice fed with a high-fat diet Low-fat diet (LL) and high-fat diet On the 20th week after feeding (HL) and starting PGD administration, blood was collected from each group of mice under ether anesthesia with parin into the vena cava of the lower limbs. After blood collection, the mice were killed by excessive ether loading and rapidly visceral. Adipose (liver, mesentery) tissue, epididymal adipose tissue, spinal subcutaneous adipose tissue were collected, and the results of measuring the weight of each tissue are shown in the table below. Using the organs thus obtained, The measurements described below were also performed.

  In addition, each value in the following table is tested by Fisher's proted LSD using Super A NOVA, and is expressed as an average value ± standard error of 7 animals in each group, p <0.05 is considered significant, and others The same was done in the test.

As is clear from the above table, the liver weight, mesenteric adipose tissue weight, epididymal adipose tissue weight, and subcutaneous adipose tissue weight were higher than those in the LL fed group. It increased by ingestion, and it was found that visceral fat obesity due to fat accumulation was induced in mice in the HF feeding group.
However, in the PGD 500 mg / kg administration group and the PGD 2000 mg / kg administration group at the same time as feeding with HF, increases in liver weight, mesenteric adipose tissue weight, epididymal adipose tissue weight, and subcutaneous adipose tissue weight were significant in all groups. Was suppressed.

(4) Measurement of neutral fat and total cholesterol level in liver Using the liver of each group of mice obtained in (3) above, clevis-ringer phosphate buffer (pH 7.5) was added, and a commercially available homogenizer was used. A 10% by weight homogenate solution of the liver was prepared.
The 10% by weight homogenate solution of each group of mouse livers prepared in this manner was extracted by adding a 20-fold amount of chloroform: methanol (2: 1) mixture, and the extract was used as a sample to obtain a neutral fat value. (TG) and total cholesterol level (TC) were measured.
The neutral fat value was measured by a triglyceride-test Wako measurement kit (acetyl-acetone method) manufactured by Wako Pure Chemical Industries, Ltd., and the total cholesterol value (TC) was measured by the ZaK method (ferric chloride-acetic acid method). . The results obtained are shown in the table below.

As is clear from the above table, it was found that the neutral fat in the liver of mice in the HF fed group was elevated and the visceral fat was accumulated and became visceral obese as compared with the LL fed group. .
However, it was found that even when HF was consumed, visceral fat accumulation was significantly suppressed by administration of PGD (500 mg / kg, 2000 mg / kg).

  The increase in the total cholesterol level in the liver was observed more strongly in the LL feeding group than in the HF feeding group, but the administration of PGD (500 mg / kg and 2000 mg / kg) also caused the accumulation of total cholesterol in the liver. It was found that it was significantly suppressed.

(5) Measurement of sucrose-degrading enzyme activity in the small intestine Generally, sugar is hardly absorbed from the small intestine unless it becomes a monosaccharide head. Therefore, digestion of the diet progresses, starch and polysaccharides are decomposed to disaccharides, and when they reach the small intestine, the small intestine secretes disaccharide degrading enzymes, for example, sucrose degrading enzymes, and enzymatic decomposition of sucrose promotes absorption in the body as glucose It is known that

Accordingly, the sucrose degrading enzyme activity in the small intestinal mucosa was measured using the small intestine of each group of mice obtained in (3) above.
After removing the contents of the small intestine, it was washed with physiological saline, the mucous membrane was peeled off from the small intestine, clevis-ringer phosphate buffer (pH 7.5) was added, and a 10% by weight homogenate of the small intestinal mucosa using a commercially available homogenizer. A liquid was prepared.
The sucrose-degrading enzyme activity of the small intestinal mucosal enzyme was measured using the 10% by weight homogenate solution of the mouse small intestinal mucosa of each group thus prepared. That is, 0.5 ml of clevis-Ringer buffer containing 10 mM sucrose was added to 0.5 ml of 10% by weight homogenate solution of small intestinal mucosa obtained from each group of mice, and the mixture was heated and shaken at 37 ° C. for 30 minutes. The reaction was stopped by heating at 45 ° C. for 45 seconds to deactivate the sucrose-degrading enzyme, and the concentration of the produced glucose was measured with a glucose B-test measurement kit manufactured by Wako Pure Chemical Industries. Degradative enzyme activity was measured. The results are shown in the following table.

As is apparent from the above results, in the small intestine of mice in the PGD administration group together with HF, the PGD (500 mg / kg) administration group and the PGD (2000 mg / kg) administration group were compared with the LL feeding group and the HF feeding group. In any of the administration groups, the amount of glucose produced was remarkably low, and it was found that the activity of sucrose degrading enzyme was suppressed.
Therefore, the phenylglucoside derivative (PGD) according to the present invention significantly suppresses the production of glucose from sucrose in the small intestine, controls the absorption of excess glucose from the small intestine causing obesity and diabetes, and improves and prevents obesity. It is thought that it is useful.

In the above results, the LL feeding group showed an increase in sucrose degrading enzyme activity compared to the HF feeding group, whereas HF contained 17.1% of corn starch in the diet as starch. Since LL is higher in concentration and contains 41.5% corn starch, it is considered to be a phenomenon caused by an increase in sucrose degrading enzyme activity in the small intestine of mice in the LL feeding group during long-term breeding. It is done.
Therefore, from the test results (3) to (5) above, it is clear that the phenylglucoside derivative according to the present invention is effective in preventing and improving visceral obesity.

(6) Measurement of blood lipids The mice of the above 4 groups were fed with a low fat diet (LL) and a high fat diet (HL) and fasted for 5 hours at 6 weeks, 10 weeks and 15 weeks after the start of PGD administration. Later, blood was collected from the tail vein of mice, plasma was separated, and blood neutral fat (TG) and total cholesterol (TC) were measured using cholesterol E-test Wako and triglyceride E test Wako, respectively. In addition, blood triglycerides and total cholesterol were measured in the same manner for blood selected at week 20, and free fatty acids were measured using a NEFA-Test Wako measurement kit. The results are shown in the following table.

From the above table, it was found that in all cases at 6, 10, 15, and 20 weeks, the HF fed group increased blood triglycerides compared to the LL fed group.
However, it was found that in the group administered with PGD together with the HF diet, blood neutral fat was significantly reduced in a dose-dependent manner.
In particular, it was found that blood neutral fat can be more strongly reduced in the group administered with PGD (2000 mg) together with the HF diet, and blood neutral fat can be reduced than in the LL feeding group.

From the above table, it was found that in all cases at 6, 10, 15, and 20 weeks, the HF fed group increased the blood total cholesterol level compared to the LL fed group.
However, it was found that the blood cholesterol level was significantly decreased in a dose-dependent manner in the group administered with PGD together with the HF diet.

From the above table, it was found that in the 20th week, the HF feeding group increased the blood free fatty acid level compared to the LL feeding group.
However, it was found that in the group in which PGD was administered together with the HF diet, the blood free fatty acid level was significantly decreased in a dose-dependent manner.

  In particular, it was found that the group administered with PGD (2000 mg) together with the HF diet was able to decrease the blood free fatty acid level more strongly and decreased the blood free fatty acid level than the LL feeding group.

  From the above results, it was found that the phenylglucoside derivative according to the present invention has a strong visceral obesity improvement and prevention effect.

  According to the present invention, a crude sugar-containing phenylglucoside derivative-containing colored component that has been conventionally disposed of in the production process of sucrose can be safely used as a composition for improving and preventing visceral fat obesity.

Claims (4)

  A composition for improving and preventing obesity comprising a phenylglucoside derivative as an active ingredient and a pharmaceutically acceptable excipient.   The composition according to claim 1, wherein the phenylglucoside derivative is derived from crude sugar.   The composition according to claim 1, wherein the phenylglucoside derivative is 3,4-dimethoxyphenyl-β-D-glucoside.   A health food comprising the composition according to claim 1 added thereto.