JP2000247880A - Fat decomposition-accelerating agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject accelerating agent exhibiting an antiobestic effect and having a high safety by containing a specific compound isolated from a plant such as Satsuma mandarin to accelerate the decomposition of fat in fat cells. SOLUTION: This accelerating agent contains a compound of the formula (Me is methyl). The compound of the formula is obtained by extracting it from Satsuma mandarin, (Citrus unshiu), bitter orange (Citrus aurantium), wild orange (Citrus tachibana), etc., of plants belonging to Rutaceae, and separating and purifying by means of a silica column chromatography. Further, it is preferable to perform the above extraction by using peels of the fruit of the above plants, and as the extracting solvent, water or a lower alcohol is preferably used and the extraction is preferably performed at 0-120 deg.C. The above accelerating agent is preferably used by preparing as a form of a cosmetic or a food. Also, it is preferable to contain 0.0001-99 wt.% compound of the formula in the objective accelerating agent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a lipolysis accelerator containing a highly safe compound represented by the following formula (1) as an active ingredient. The lipolysis accelerator of the present invention has an excellent anti-obesity effect, and can be widely used in various fields such as cosmetics, foods, and pharmaceuticals.

[0002]

2. Description of the Related Art In recent years, Japanese meals have become remarkably westernized,
High calorie is progressing. In particular, excessive intake of lipids causes obesity, which is also called modern civilization's disease. Obesity is one of social problems because it is closely related to various diseases such as hyperlipidemia, arteriosclerosis, and diabetes. In addition, the desire to reduce the aesthetics of Japanese people in recent years has become extremely strong.

However, to date, none of the skin cosmetics marketed for anti-obesity has been found to be extremely effective. On the other hand, in the literature, for example, an example using an adrenergic β-stimulant such as epinephrine or theophylline (US Pat. No. 4,525,359) is described, but these have problems in safety.

[0004] Obesity treatments generally include diet, exercise and pharmacotherapy. Of these, diet and exercise are usually combined to treat obesity.
However, these therapies take a long time to take effect. Therefore, it requires strong will, but is extremely difficult to implement in a busy world. On the other hand, in drug therapy, appetite suppressants such as mazindol and fenfluramine have been developed, but are known to have side effects such as dry mouth and depression.

[0005]

An object of the present invention is to provide a highly safe lipolysis promoter which exerts an antiobesity effect by promoting the decomposition of fat in fat cells.

[0006]

Means for Solving the Problems The present inventors screened various natural products and compounds in order to solve the above problems. This screening was carried out using, as an index, the effect of decomposing fat in free adipocytes obtained from rat epididymal adipose tissue using collagenase. As a result, they have found that the compound represented by the following formula (1) has the intended effect, and have completed the present invention.

That is, the present invention provides the following formula (1):

Embedded image It is intended to provide a lipolysis promoter containing a compound represented by the formula: wherein Me is methyl. In a preferred embodiment of the present invention, the lipolysis promoter is prepared in the form of a cosmetic or food.

[0008]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail. The compound (1) used in the present invention is a known substance, and is a mouse leukemia cell (M1 cell) and a human leukemia cell.
(HL-60 cells) is known to have a differentiation inducing effect [Chem. Pharm. Bull. 41 (4), 714-719, 19
93]. However, it has not been known that this compound has a lipolysis promoting effect. This compound (1) can be produced by chemical synthesis, but is usually obtained by isolating from a plant containing the compound (1).

Compound (1) is used as a citrus plant, Citrus unshiu or Citrus auranti
um), or by extraction from Citrus tachibana.

When compound (1) is prepared by extraction from a plant, the plant parts used include leaves, stems, bark,
Examples include flowers, fruits, and pericarp, but pericarp is particularly preferred. The plant may be used as it is after collection, or may be used after drying.

As the solvent used for the extraction, water or one or more kinds of organic solvents such as lower alcohols (eg, methyl alcohol, ethyl alcohol or butanol), acetone and ethyl acetate are appropriately mixed and used. it can. Preferred extraction solvents are water or lower alcohols alone or a mixture of water and lower alcohols.

[0012] The extraction may be performed in a usual manner. The extraction temperature is usually in the range of 0 to 120C, preferably 40 to 120C. Although the extraction time varies depending on the extraction temperature, it is generally 1 to 10 days for extraction near room temperature, and 0.5 to 120 hours for extraction at 40 ° C. or higher.

The compound (1) can be separated and purified from the plant extract thus obtained by a method such as silica column chromatography or preparative HPLC. Compound (1) thus separated and purified
Can be used as an active ingredient of the lipolysis promoter of the present invention.

The lipolysis accelerator of the present invention can be prepared in any form for internal use or external use. For internal use, the lipolysis promoter of the present invention is prepared in the form of a food or a pharmaceutical. In the case of external use, the lipolysis promoter of the present invention is prepared in the form of cosmetics, quasi-drugs, or pharmaceuticals.

The lipolysis promoter of the present invention is preferably prepared and used in the form of a cosmetic or food. Examples of the form of the cosmetic include a gel cream, a face wash cream, a lotion, a milky lotion, a pack and the like, and other components other than the active ingredient are selected according to the form. Examples of the form of the food include granules, tablet confectionery, jelly, candy, and beverages, and other ingredients other than the active ingredient are selected according to the form.

The compounding ratio of the compound (1) in the lipolysis accelerator of the present invention varies depending on the form of the lipolysis accelerator.
Usually, 0.0001 to 99 relative to the total amount of the lipolysis accelerator
% By weight, preferably 0.001 to 50% by weight, more preferably 0.001 to 20% by weight, and still more preferably 0.00% by weight.
It is in the range of 1 to 10% by weight.

The lipolysis accelerator of the present invention comprises, in addition to the compound (1) which is an active ingredient, various components generally used in preparing them according to the form of various lipolysis accelerators. contains. Other components used in preparing cosmetics include, for example, oils, surfactants, humectants, thickeners, preservatives, fragrances, colorings, drugs, and the like. The cosmetics of the present invention can include one or more of these components. Also, other ingredients used in preparing foods,
Food ingredients commonly used in this field may be used, and examples of these food ingredients include lactose, dextrose, sucrose, sorbitol, mannitol, apple fiber, soy fiber, meat extract, black vinegar extract,
Examples include gelatin, corn starch, honey, animal and vegetable fats and oils, and polysaccharides. The food product of the present invention can include one or more of these food ingredients.

The food of the present invention further comprises one or more ingredients such as a lubricant, an emulsifier, a suspending agent, an antioxidant, a preservative, a sweetener and a flavoring agent in addition to the above-mentioned food ingredients. be able to. Further, it may further contain other active ingredients (including water-soluble vitamins and oil-soluble vitamins). Those skilled in the art can easily select appropriate components that do not interfere with the lipolysis-promoting effect of compound (1).

The lipolysis promoter of the present invention may be produced by a method well known in the art depending on its form. In addition, the lipolysis promoter of the present invention can be appropriately applied or ingested according to the form.

[0020]

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

Example 1 Preparation of Compound (1) 3 L of 50% (v / v) ethanol was added to 1 kg of spruce (purchased from Takasago Pharmaceutical Co., Ltd.) and extracted at 40 ° C. for 2 days. The mixture was filtered to obtain 2.1 L of filtrate. The filtrate was concentrated under reduced pressure at a temperature of 60 ° C. or lower, and the obtained concentrated liquid was separated with ethyl acetate / distilled water. The ethyl acetate layer was collected and dried over anhydrous sodium sulfate. Next, the ethyl acetate layer was concentrated under reduced pressure at 60 ° C. or lower to obtain an oily substance. This oily substance was subjected to silica column chromatography [elution solvent: ethyl acetate: n-hexane = 1: 1 (750 ml) → 2: 1 (1050 ml) → 5: 1 (780 m
l)] to give a fraction containing compound (1). Compound (1) was mainly contained in the fraction eluted with ethyl acetate: n-hexane = 2: 1, and the fraction containing compound (1) was confirmed by thin-layer chromatography (developing solvent: ethyl acetate). . Subsequently, the fractions containing the compound (1) were combined and purified by silica column chromatography (elution solvent: chloroform: methanol = 50: 1) to obtain 142 mg of the compound (1).

The measurement results of the proton nuclear magnetic resonance spectrum of the compound (1) were as follows. ¹ H-NMR (CDCl ₃ , 500 MHz) δ (ppm): 3.77 (s, 3H), 3.8
3 (s, 3H), 3.84 (s, 3H), 3.87 (s, 3H), 3.96 (s, 3H), 4.01
(s, 3H), 6.85 (s, 1H), 7.15 (d, J = 8.6Hz, 1H), 7.53 (d, J =
2.1Hz, 1H), 7.64 (dd, J = 2.1, 8.6Hz, 1H). This result is shown in the literature [Chem. Pharm. Bull., 35 (7), 3025-302.
8, 1987], which is in good agreement with the data of compound (1). Further, the purity of the compound (1) was analyzed by high performance liquid chromatography (HPLC) under the following conditions. As a result, the chemical purity was 99.2%. HPLC analysis conditions : Column: μBONDASPHERE C18, 3.9 × 150 mm (manufactured by Waters) Fluidized bed: acetonitrile (in 2% acetic acid aqueous solution) 15-60
%, 30 minutes Flow rate: 1.0 mL / min Column temperature: 40 ° C. Detection: UV 280 nm Retention time: 29.05 minutes

Example 2 Measurement of Lipolytic Activity on Rat Epididymal Adipocytes It is known that glycerol and free fatty acids are released when fat in adipocytes is degraded. Lipolytic activity was measured by quantifying the free fatty acids released into the medium. Quantification of free fatty acids was performed by an enzymatic method. According to the method of rodbell [Rodbell, M, J. Biol. Chem., 239, 375 (1964)], free adipocytes were obtained from epididymal adipose tissue of four male Wistar 8-week-old rats using collagenase solution. Prepared. Krebs Ringer (Kreus Ringer) containing bovine serum albumin prepared so that the concentration of compound (1) prepared in Example 1 becomes 10 μg / mL and 50 μg / mL.
bs Ringer) 37 cells in bicarbonate buffer
After incubation at 90 ° C for 90 minutes, the released fatty acids were measured using a commercially available kit (Wako Pure Chemical Industries, NEFA C-Test Wako). The lipolytic activity was compared with the control without compound (1). Table 1 shows these results.
Shown in

[Table 1] As is clear from Table 1, the concentration of the compound (1) was 10 μg.
/ ML and 50 μg / mL, the amount of free fatty acids from adipocytes was 110.
65% and 219.91%.

Example 3 Measurement of lipolytic activity on rat subcutaneous fat cells The abdominal skin tissue of 5 Wistar 8-week-old male rats was exfoliated together with the subcutaneous adipose tissue to a diameter of 3 cm and a Franz type 1.8 cm diameter was obtained. Set in diffusion cell. Compound (1) prepared in Example 1 and white petrolatum were added at 1.0: 99.
0 and 2.0: 98.0 in a weight ratio of this mixture
(0.2 g) was evenly applied to the rat skin surface, and the lower cell was filled with Krebs-Ringer bicarbonate buffer. After leaving at 37 ° C. for 5 hours, the glycerol released in the buffer solution in the lower cell was replaced with F-kit glycerol.
(Boehringer Mannheim). As a control, 100% petrolatum was used. The results obtained are shown in Table 2 below. Results are shown as% ± SD% relative to control (n = 4).
Displayed with.

[Table 2] The results in Table 2 revealed that compound (1) acts on subcutaneous adipocytes through the skin and promotes release of glycerol from adipocytes. That is, it was revealed that the compound (1) has a lipolysis promoting activity.

Example 4 Gel Cream The compound (1) obtained in Example 1 is used as an active ingredient in the following gel cream formulation (100% by weight in total).

[Table 3] Stir and mix all components at room temperature to make a homogeneous solution,
The pH was adjusted to 6.5 to obtain a gel cream.

Comparative Example 1 Conventional Gel Cream The same gel as in Example 4 except that the compound (1) was replaced with purified water was used as a conventional gel cream.

Example 5 Lotion The compound (1) obtained in Example 1 is used as an active ingredient in the following lotion formulation (total 100% by weight).

[Table 4] Stir and mix all components at room temperature to make a homogeneous solution,
The pH was adjusted to 5.5 to obtain a lotion.

Example 6 Emulsion The compound (1) obtained in Example 1 is used as an active ingredient in the following emulsion formulation (total 100% by weight).

[Table 5] The component A is dissolved by heating and kept at 80 ° C. Separately, the component B heated and dissolved at 80 ° C. is added to the component A and mixed well. The mixture was cooled while stirring, and the component C was added at 50 ° C. to obtain an emulsion.

Example 7 Food A Using the compound (1) obtained in Example 1 as an active ingredient, a food A having the following composition was prepared.

[Table 6] After mixing each component in a fluidized-granulation apparatus, water was sprayed to perform granulation, and the mixture was dried at an air inlet temperature of 80 ° C.

Comparative Example 2 Comparative Food A food prepared by using dextrin instead of compound (1) in Food A of Example 7 was prepared as a comparative food.

Example 8 Food B Using the compound (1) obtained in Example 1 as an active ingredient, a food B having the following composition was prepared.

[Table 7] After sufficiently mixing the above components with 10 kg of water, the mixture was heated to 60 ° C. to form a paste. This was put into 50 kg of soybean oil which had been heated to 70 ° C. in advance, and thoroughly mixed with a stirrer equipped with squid-type stirring blades (two pieces). After confirming that the paste was dispersed in the soybean oil, the mixture was cooled to 5 ° C.
And filtered to separate the resulting particles. The obtained particles were washed with n-hexane and then dried to obtain microcapsules containing the above components.

Example 9 Tablet A tablet was prepared using the following ingredients.

[Table 8]

Example 10 Using the gel-like cream of Example 4 and the gel-like cream of Comparative Example 1, a 30-year-old female from 20 to 50 years of use test (external application, 1 day per day) After bathing twice). After use, the hips and waist circumference were measured and compared with those before the start of the test. These results are shown in Table 9 below.
And Table 10.

[Table 9]

[Table 10] As is clear from these tables, the gel cream containing the compound (1) showed an excellent anti-obesity effect.

Example 11 Using the food of Example 7 and the food of Comparative Example 2,
A two-month use test (2.0 g after each meal; three times a day) was performed on 15 men and women aged 5 to 50 years each. After use, body weight was measured and compared to before the start of the test. The results are shown in Tables 11 and 12 below.

[Table 11]

[Table 12] As is clear from these tables, the food containing compound (1) exhibited an excellent anti-obesity effect.

[0035]

From the above results, it has been clarified that the compound (1) shows an excellent lipolytic activity, and the lipolysis accelerator containing the compound (1) shows an excellent anti-obesity effect. That is, the lipolysis promoter of the present invention is intended for anti-obesity,
It can be widely used in the form of cosmetics, foods, pharmaceuticals, etc.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification FI FI Theme Court ゛ (Reference) // C07D 311/30 C07D 311/30 (72) Inventor Kunio Kosaka 2-2-2 Muroya, Nishi-ku, Kobe-shi, Hyogo Prefecture No. 3 Nagase & Co., Ltd. R & D Center F-term (reference) 4B018 MD07 MD31 MD36 MD45 MD52 MD58 MD73 MD77 MD92 ME14 MF01 4C062 EE49 4C083 AA112 AA122 AB032 AC022 AC072 AC122 AC132 AC302 AC432 AC442 AC482 AC841 AC842 AC852 AD092 AD222 AD512 AD662 CC04 CC05 DD41 EE11 EE50 4C086 AA01 AA02 BA08 MA01 NA14 ZA70 ZC33

Claims (3)

[Claims] 1. The following formula (1): [Wherein Me is methyl] a lipolysis accelerator comprising, as an active ingredient, a compound represented by the formula: 2. The lipolysis accelerator according to claim 1, which is prepared in the form of a cosmetic. 3. The lipolysis accelerator according to claim 1, which is prepared in the form of a food.