JP2007186427A - Fat cell differentiation inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fat cell differentiation inhibitor that can be daily taken continuously, is safe and sufficiently suppresses fat accumulation of precursor fat cells. <P>SOLUTION: The fat cell differentiation inhibitor comprises an embryo of a lotus and/or extracts of yacon. The fat cell differentiation inhibitor suppresses fat accumulation of precursor fat cells. Foods, quasi-drugs and pharmaceuticals containing the embryo of a lotus and/or extracts of yacon exhibit fat cell differentiation-inhibiting actions and are effective for prevention and amelioration of obesity and diabetes. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a novel medicinal ingredient having an action of inhibiting differentiation of preadipocytes and a pharmaceutical, a quasi-drug, a cosmetic and a food containing the same as an active ingredient. This medicinal component is mainly contained in lotus germ and yacon.

  Adipocytes are cells that originate from mesodermal pluripotent stem cells and differentiate into mature adipocytes via lipoblasts, preadipocytes, and immature adipocytes. White adipocytes are responsible for fat synthesis, storage, and release. And brown adipocytes responsible for heat production by the action of mitochondrial uncoupling protein (UCP).

  Obesity refers to a state in which neutral fat is excessively accumulated in white adipocytes. Therefore, obesity is caused by the excess of calories in white fat cells due to excess calories consumed due to physical factors, dietary factors, mental factors, central factors, metabolic factors, and lack of exercise. Invited after being converted to. However, in recent years, it has become clear that the number of white adipocytes increases even after adulthood, and this has been found to further promote obesity. Therefore, suppressing differentiation from precursor white fat cells to mature white fat cells and reducing the number of mature white fat cells is expected to lead to improvement in obesity.

  There are not many drugs, quasi-drugs, cosmetics and foods that suppress the increase in the number of fat cells and show an anti-obesity effect. There exists what uses activated whey as an active ingredient (patent document 2). There is also an attempt to apply ω-3 highly unsaturated fatty acid as an active ingredient to an external preparation for skin (Patent Document 3). However, there is still no sufficient effect, and a novel preadipocyte differentiation inhibitor has been desired.

Japanese Unexamined Patent Publication No. 6-293396 JP 2000-37738 A Japanese Patent Laid-Open No. 11-130656

  An object of the present invention is to provide an effective and highly safe preadipocyte differentiation inhibitor, an anti-obesity agent, and a medicine and food containing them.

  The present inventor performed screening using mouse preadipocytes 3T3-L1 with the suppression of differentiation into adipocytes as an index. As a result, it was found that lotus germ extract or yacon extract suppresses differentiation from preadipocytes to adipocytes, and further, combined use of lotus germ and yacon extracts, synergistic differentiation suppression effect As a result, the present invention has been completed.

  The lotus germ used in the present invention is a water lily family germ, for example, a lotus of the genus Lotus (Nelumbo nuifera Gaertn.), N. lutea Pers. Embryos such as Victoria regia Lindl. Can be used. Examples of products distributed as crude drugs include lotus wicks, lotus wicks, etc., which can be purchased and used. Various parts of lotus are used for medicinal purposes, and each has a different medicinal effect. For example, lotus leaves (carcasses) of the lotus genus have a different medicinal effect against diarrhea, hemostasis, fruits (lotus meat) and stamens (lotus), and germs (lotus core, lotus core) different from antipyretic. ing. In addition, a large amount of starch and sugar raffinose are known as fruit components, whereas leaves contain alkaloids such as loemerin and luciferin, and germs contain different alkaloids such as methylcolyparin and rotsin. It is known (primary color Japanese medicinal plant pictorial book, Koichi Kimura, Jun Kimura, nursery company).

  The lotus germ extract used in the present invention can be extracted from a lotus germ with a solvent. The extraction method is not particularly limited, and for example, it may be a heat extraction or a room temperature extraction.

  In the present invention, a yacon is a plant belonging to the family Asteraceae that is native to Peru, and its scientific name is Polymnioa sonchifolia.

  The extract of yacon used in the present invention is obtained by extracting from a part or all of a plant such as leaves, stems, flowers, seeds, fruits, rhizomes and roots of yacon. Preferably, it is obtained by extraction from one or both of leaves and stems. The preparation method is not particularly limited, and for example, it may be extracted by heating or may be extracted at room temperature.

  Examples of the solvent to be extracted include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol). ), Ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.) Can be mentioned. Preferable are polar solvents such as water and lower alcohols, and particularly preferable are water and ethanol. These solvents may be used alone or in combination of two or more.

  The above extract may be used as it is, or may be used after treatment such as concentration, dilution, filtration, decolorization with activated carbon, deodorization, ethanol precipitation or the like, if necessary. Furthermore, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.

  As the adipocyte differentiation inhibitor of the present invention, the above extract may be used as it is, and a diluent may be used as long as the effect of the extract is not impaired. The ratio when the lotus germ extract and the yacon extract are blended with the adipocyte differentiation inhibitor is not particularly limited. The diluent may be a solid, liquid, or semi-solid, and examples include the following. That is, excipients, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, dispersants, buffers, fragrances, preservatives, solubilizers, solvents, and the like. Specifically, lactose, sucrose, sorbit, mannitol, starch, precipitated calcium carbonate, heavy magnesium oxide, talc, calcium stearate, magnesium stearate, cellulose or derivatives thereof, amylopectin, polyvinyl alcohol, gelatin, surface activity Agents, water, physiological saline, ethanol, glycerin, propylene glycol, cacao butter, lauric fat, petrolatum, paraffin, higher alcohol and the like.

  The adipocyte differentiation inhibitor of the present invention can be used in any of foods, cosmetics, quasi drugs, and pharmaceuticals. Examples of the dosage form include powders, granules, tablets, sugar-coated tablets for oral use, Capsules, syrups, pills, suspensions, solutions, emulsions and the like can be mentioned. Examples of parenteral use include dosage forms such as lotions, creams, emulsions, and bath preparations. Moreover, it can also be set as an injection and a suppository.

  The intake of the extract used in the present invention varies depending on the dosage form, purpose of use, age, body weight, etc., but is usually 0.001% by weight or more, preferably 0.01- 50.0% by weight is preferable. In addition, the method for adding medicinal ingredients in the formulation may be added in advance or during production, and may be appropriately selected in consideration of workability. The daily dose of the lotus germ extract is 1-1000 mg, preferably 5-200 mg. Yacon extract is 1-1000 mg, preferably 5-500 mg. Since the administration method and dosage vary depending on various conditions, an amount smaller than the above-mentioned administration range may be sufficient, or it may be necessary to administer beyond the range.

  The lotus germ or yacon extract of the present invention suppresses the differentiation of preadipocytes, and thus suppresses fat accumulation, and is effective in preventing and improving obesity and diabetes.

  In order to describe the present invention in detail, examples of production of the extract used in the present invention, formulation examples of the present invention, and experimental examples are given as examples, but the present invention is not limited thereto. The part of the amount shown in the examples indicates part by weight, and% indicates% by weight.

Production Example 1 Hot water extract of lotus embryos 100 mL of purified water was added to 10 g of dried lotus germ (lotus core), extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated and freeze-dried. As a result, 2.6 g of hot water extract was obtained.

Production Example 2 50% ethanol extract of lotus germ 50 ml of purified water and 50 mL of ethanol were added to 10 g of dried lotus germ (lotus wick), extracted at room temperature for 3 days, filtered, and the filtrate was concentrated and lyophilized. As a result, 2.1 g of 50% ethanol extract was obtained.

Production Example 3 Ethanol extract of lotus germ 100 g of ethanol was added to 10 g of dried lotus germ (lotus wick), extracted at room temperature for 3 days, filtered, the filtrate was concentrated and freeze-dried to obtain an ethanol extract. 0.4 g was obtained.

Production Example 4 50% 1,3-butylene glycol extract of lotus germ 50 g of purified water and 50 g of 1,3-butylene glycol (1,3-BG) are added to 10 g of dried lotus germ (lotus wick) and at room temperature. Extraction was performed for 7 days, followed by filtration to obtain 90 g of 50% 1,3-BG extract.

Production Example 5 Hot Water Extract of Yacon 1 kg of purified water was added to 40 g of a dried yacon stem and leaf mixture (made in China) and extracted at 95-100 ° C. for 2 hours. The obtained extract was concentrated and dried to obtain 11 g of a hot water extract of yacon.

Production Example 6 1,3-butylene glycol extract of yacon 500 g of purified water and 500 g of 1,3-butylene glycol were added to 100 g of dried dried yacon stems and leaves (from China), and extracted at room temperature for 2 weeks. . After extraction, the mixture was filtered to obtain 950 g of a 50% 1,3-butylene glycol extract of Yacon.

Production Example 7 Propylene glycol extract of yacon 500 g of purified water and 500 g of propylene glycol were added to 30 g of dried yacon whole plant (made in China) and extracted at room temperature for 2 weeks. After extraction, the mixture was filtered to obtain 950 g of a yacon propylene glycol extract.

  The extract of the present invention can be formulated as the following formulation examples.

Formulation Example 1 Lotion Formulation Formulation 1. 50% 1,3-butylene glycol extract of lotus germ (Production Example 4) 0.05 part Yacon 1,3-butylene glycol extract (Production Example 6) 0.05
3. 1,3-butylene glycol 8.0
4). Glycerin 2.0
5). Xanthan gum 0.02
6). Citric acid 0.01
7). Sodium citrate 0.1
8). Ethanol 5.0
9. Pantothenic acid ethyl alcohol 0.1
10. Methyl paraoxybenzoate 0.1
11. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
12 Fragrance 0.1
13. Purified water 84.37
[Production method] Components 1 to 7 and 13 and components 8 to 12 are uniformly dissolved, and both are mixed and filtered to obtain a product.

Comparative Formulation Example 1 Conventional Lotion In Formulation Example 1, 50% 1,3-butylene glycol extract of lotus germ and 1,3-butylene glycol extract of Yacon were replaced with purified water to obtain a conventional lotion.

Formulation Example 2 Cream Formulation Formulation 1. 1. 50% ethanol extract of lotus germ (Production Example 2) 0.03 part Yacon propylene glycol extract (Production Example 7) 0.07
3. Squalane 5.5
4). Olive oil 3.0
5). Stearic acid 2.0
6). Beeswax 2.0
7). Octyldodecyl myristate 3.5
8). Polyoxyethylene cetyl ether (20E.O.) 3.0
9. Behenyl alcohol 1.5
10. Glycerol monostearate2.5
11. Fragrance 0.1
12 Methyl paraoxybenzoate 0.2
13. Ethyl paraoxybenzoate 0.05
14.1,3-Butylene glycol 8.5
15. Purified water 68.05
[Manufacturing method] Components 3 to 10 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 2 and ingredients 12 to 15 are dissolved by heating and mixed, and kept at 75 ° C. to obtain an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. The component 11 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Comparative Formulation Example 2 Conventional Cream In Formulation Example 2, a 50% ethanol extract of lotus germ and a propylene glycol extract of Yacon were replaced with purified water to obtain a conventional cream.

Formulation Example 3 Emulsion Formulation Formulation 1. Hot water extract of lotus germ (Production Example 1) 0.005 part2. Yacon hot water extract (Production Example 5) 0.005
3. Squalane 5.0
4). Olive oil 5.0
5). Jojoba oil 5.0
6). Cetanol 1.5
7). Glycerol monostearate 2.0
8). Polyoxyethylene cetyl ether (20E.O.) 3.0
9. Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
10. Fragrance 0.1
11. Propylene glycol 1.0
12 Glycerin 2.0
13. Methyl paraoxybenzoate 0.2
14 Purified water 73.19
[Manufacturing method] Components 3 to 9 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1, 2 and 11-14 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. The component 10 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Formulation Example 4 Bath preparation formulation Lotus extract ethanol extract (Production Example 3) 4.9 parts2. Yacon 1,3-butylene glycol extract (Production Example 6) 0.1
3. Sodium bicarbonate 50.0
4). Menthol 0.1
5). Yellow No. 202 (1) 0.01
6). Fragrance 0.1
7). Sodium sulfate 44.79
[Production Method] Components 1 to 7 are uniformly mixed to obtain a product.

Formulation Example 5 Powder formulation Formulation amount 1. Hot water extract of lotus germ (Production Example 1) 15 parts2. Yacon hot water extract (Production Example 5) 5
3. Dried corn starch 30
4). Microcrystalline cellulose 50
[Manufacturing method] Components 1 to 4 are mixed to obtain a powder.

Formulation Example 6 Tablet formulation Formulation amount 1. 50% ethanol extract of lotus germ (Production Example 2) 3 parts Yacon hot water extract (Production Example 5) 2
3. Dried corn starch 25
4). Carboxymethylcellulose calcium 20
5). Microcrystalline cellulose 40
6). Polyvinylpyrrolidone 7
7). Talc 3
[Production Method] Components 1 to 6 are mixed, then 10% water is added as a binder, and after extrusion granulation, it is dried. Ingredient 7 is added to the formed granules, mixed and compressed into tablets. One tablet is 0.52 g.

Comparative Formulation Example 3 Conventional Tablet In Formulation Example 6, a 50% ethanol extract of lotus germ and a hot water extract of Yacon were replaced with dry corn starch to obtain a conventional tablet.

Formulation Example 7 Tablet Confectionery Formulation 1. 1. 50 parts ethanol extract of lotus germ (Production Example 2) 1.0 part Yacon 1,3-butylene glycol extract (Production Example 6) 0.5
3. Dried corn starch 50.0
4). Erythritol 40.0
5). Citric acid 5.0
6). Sucrose fatty acid ester 3.4
7). Fragrance 0.1
[Manufacturing method] Components 1 to 5 are mixed, and 10% water is added as a binder to perform fluidized bed granulation. Ingredients 6 and 7 are added to the formed granules, mixed and compressed into tablets. One tablet is 1.0 g.

Formulation Example 8 Beverage Formulation Amount 1. Ethanol extract of lotus germ (Production Example 3) 2.5 parts Yacon propylene glycol extract (Production Example 7) 1.5
3. Stevia 0.05
4). Malic acid 5.0
5). Fragrance 0.1
6). Water 90.85
[Manufacturing method] Components 1 to 5 are stirred and dissolved in a part of component 6 water. The remaining water of component 6 is then added and mixed.

Comparative Formulation Example 4 Conventional Beverage In Formulation Example 8, the ethanol extract of lotus germ and the propylene glycol extract of yacon were replaced with water to obtain a conventional beverage.

Effect of inhibiting differentiation into adipocytes by insulin stimulation using mouse preadipocytes Mouse preadipocytes (3T3-L1) show a fibroblast-like morphology under normal culture conditions. However, after making it confluent, by applying insulin stimulation, fat particles are formed in the cells and differentiated into fat cells. Using this differentiation-inducing trait, it becomes possible to search for a differentiation inhibitor for adipocytes. The differentiation inhibitor obtained by this search can suppress obesity, which is one of the causes of diabetes.
In a 35 mm dish, mouse preadipocytes (3T3-L1) were cultured in 2 mL of culture medium (DMEM medium containing 10% fetal calf serum (Dulbecco's modified Eagle medium)) in the presence of 5% CO ₂ until reaching confluence. Cultured at 37 ° C. After the cells reached confluence, the medium was removed by aspiration, and the cells were washed with phosphate buffered saline (hereinafter referred to as PBS), and then differentiation induction medium 1 (10% fetal bovine serum, 1 μM dexamethasone, 10 μg / mL) 2 mL of DMEM medium containing insulin, 0.5 mM 3-isobutyl-1-methylxanthine and 100 μM ascorbic acid was added. Next, a sample solution (2 to 4 μL) dissolved in DMSO was added, and the cells were cultured at 37 ° C. for 3 days in the presence of 5% CO ₂ . As a control, only DMSO was added to the differentiation induction medium. Next, the medium was aspirated and the cells were washed with PBS, and then the medium was converted to 2 mL of differentiation induction medium 2 (DMEM medium containing 10% fetal bovine serum, 10 μg / mL insulin and 100 μM ascorbic acid). The sample solution was added again and cultured for 3 days. Thereafter, the medium was removed by aspiration, the cells were washed with PBS, 2 mL of DMEM medium containing 10% fetal bovine serum and a sample solution having the same concentration were added, and further cultured for 3 days.
Nine days after the start of differentiation induction, the medium was removed by suction, washed with PBS, glutaraldehyde was added to fix the cells, and fat particles accumulated in the cells were stained with Sudan II. The stained fat granules were eluted with isopropanol containing 4% Triton X-100, and the absorbance at 490 nm was measured to evaluate the effect of inhibiting adipocyte differentiation.

The adipocyte differentiation inhibitory effect was expressed by fat accumulation rate. That is, the lower the fat accumulation rate, the higher the differentiation suppressing effect. The fat accumulation rate was calculated by the following formula.
Fat accumulation rate (%) = (absorbance at 490 nm when sample solution is added) / (absorbance at 490 nm when DMSO is added) × 100

  As a result of examining the effect of inhibiting adipocyte differentiation using the above experimental method, as shown in Table 1, the 50% ethanol extract of the lotus germ and the hot water extract of Yacon used in the present invention are concentration-dependent, respectively. Inhibited fat accumulation. Moreover, when both were used together, fat accumulation was suppressed synergistically. From the above results, it has been found that the lotus germ extract and the yacon extract each independently inhibit differentiation into adipocytes, and when used together, synergistically inhibits differentiation into adipocytes.

Usage test 1
One month use for 30 women (21 to 46 years old) using the lotion of Formulation Example 1, the cream of Formulation Example 2, the conventional lotion of Comparative Formulation Example 1 and the conventional cream of Comparative Formulation Example 2. A test was conducted. After use, the slimming effect on the feeling of skin tightening, skin tension and skin elasticity was determined by questionnaire.

  The test results are shown in Table 2. As a result, the lotion of Formulation Example 1 and the cream of Formulation Example 2 containing the lotus germ extract and the yacon extract showed an excellent slimming action as compared with the conventional lotion and the conventional cream. During the test period, there was no skin problem and there was no problem with safety. There was also no problem with the deterioration of the prescription ingredients.

Usage test 2
One month for 20 mildly obese men (35 to 55 years old) using the tablets of Formulation Example 6, the beverage of Formulation Example 8, the conventional tablet of Comparative Formulation Example 3, and the conventional beverage of Comparative Formulation Example 4. A usage test was conducted. Before and after use, body weight, subcutaneous fat thickness and waist circumference were measured to determine the slimming effect.

  These test results are shown in Table 3. As a result, the tablets of Formulation Example 6 and the beverage of Formulation Example 8 containing the lotus germ extract and the yacon extract showed an excellent slimming effect compared to the conventional tablets and the conventional beverages. During the test period, there was no subject who was unwell, and there was no problem in safety. There was also no problem with the deterioration of the prescription ingredients.

The adipocyte differentiation inhibitor, which contains lotus germ and yacon extract, has the effect of suppressing the fat accumulation of preadipocytes, so it can be incorporated into foods, quasi drugs or pharmaceuticals, etc. It is effective for prevention and improvement of obesity and diabetes.

Claims (3)

  A preadipocyte differentiation inhibitor comprising a lotus germ and / or yacon extract.   An antiobesity agent comprising the preadipocyte differentiation inhibitor according to claim 1 as an active ingredient. A pharmaceutical, quasi-drug, cosmetic and food containing the preadipocyte differentiation inhibitor according to claim 1 as an active ingredient for the purpose of preventing or improving obesity and diabetes.