JP2011201785A - Antiobestic agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antiobestic agent, a lipolysis promoter, a blood neutral fat increase inhibitor and an adiponectin production promoter that can be daily administered continuously and is effective and highly safe.SOLUTION: The antiobestic agent and the like comprise extracts of an eyebright. Though a variety of preparation forms can be selected, an internal medicine is particularly preferable. The dosage depends on the purpose of use, age, body weight, symptoms, therapeutic effects and the like, and is 0.02-500 mg/kg body weight, particularly preferably 0.1-300 mg/kg body weight. The agents act on obesity multilaterally to exhibit excellent improving effects, and are therefore useful for pharmaceuticals, quasi-drugs, foods, cosmetics and the like used for the purpose of prevention and amelioration of obesity and life style-related diseases associated with obesity.

Description

  The present invention relates to an anti-obesity agent, a lipolysis promoter, a blood neutral fat increase inhibitor, and an adiponectin production promoter characterized by containing an eyebright extract.

  In recent years, in Japan, obesity due to excessive intake of calories and lack of exercise due to a rich diet has increased rapidly. Obesity is a state in which neutral fat is excessively accumulated in fat cells. The main causes of obesity are excessive calorie intake and lack of exercise as described above, but many other factors such as constitutional factors, metabolic factors, dietary factors, mental factors, and central factors are involved. Various methods for preventing and improving obesity have been proposed from various fields.

  Representative anti-obesity agents include those that prevent and improve obesity by preventing digestion and absorption of carbohydrates and lipids. For example, one or two or more selected from the group consisting of an α-amylase inhibitor (Patent Document 1) characterized by containing okogi or an extract thereof, Hakutou, Mitsumouka and the like. It is a fat absorption inhibitor (Patent Document 2).

  A lipolysis promoter has been proposed as an agent that actively decomposes neutral fat in fat cells to prevent and improve obesity. For example, a lipolysis promoter (Patent Document 3) containing an extract of the genus Raffle genus plant as an active ingredient (Patent Document 3), a lipolysis promoter (Patent Document 4) consisting of pericarps, leaves or extracts thereof .

In general, a state where the value of neutral fat in the blood is 150 mg / dL or more is called dyslipidemia. As dyslipidemia progresses, it is known that neutral fat deposits on the blood vessel wall and causes arteriosclerosis, and further leads to serious diseases such as ischemic heart disease and cerebrovascular disorder. In addition, triglycerides rapidly absorbed in the body are released into the blood vessels, and most of them accumulate in fat cells without being converted into energy, so blood triglycerides are closely related to obesity. .
Examples of blood neutral fat elevation inhibitors include, for example, lipase inhibitors and hepatic triglyceride lowering agents (Patent Document 5) containing hesperidin and its glycosides as active ingredients, and postprandial meals containing Yunnan tea extract as active ingredients. (2) discloses a blood neutral fat elevation inhibitor (Patent Document 6).

Adiponectin is a cytokine that is specifically produced and secreted from adipocytes of adipose tissue and is closely involved in the metabolism of sugars and lipids (Non-patent Document 1). Although adiponectin is secreted from adipocytes, it is known that the blood concentration decreases due to obesity and the risk of cardiovascular disease increases (Non-patent Document 2). Adiponectin has been reported to promote fat burning by activating AMP kinase in skeletal muscle and liver (Non-patent Document 3), and is closely related to obesity. For this reason, various adiponectin production promoters have been proposed to increase the blood adiponectin concentration.
For example, an adiponectin production promoter characterized by containing an extract or processed product of nutmeg as an active ingredient (Patent Document 7), an adiponectin production promoter containing a hot water extract of Cordyceps sinensis as an active ingredient (Patent Document 8) ).

  However, the effects of the aforementioned lipolysis promoter, blood neutral fat elevation inhibitor and adiponectin production promoter are not all satisfactory, especially for obesity involving multiple factors in a complex manner. The effect of was not expected.

  The ibrite related to the present invention is a herb belonging to the family Euphorbiaceae originating in Europe, West Asia and North America, and its scientific name is Euphrasia officinalis. Ibright has been used in Europe for its own phytotherapy, and has been used as a facial wash or tea for acute and subacute inflammation of the eyes, especially conjunctivitis, blepharitis, and visual impairment. In addition, suppression of protein glycation, application to treatment of various diseases such as diabetic complications caused by glycation, retinopathy, nephropathy (Patent Document 9), suppression of IgE production to treat allergic inflammation The use to an agent (patent document 10), the use to the skin external preparation which has a whitening effect (patent document 11), etc. are proposed.

  However, there are no reports on the anti-obesity effect, lipolysis promoting effect, blood neutral fat elevation inhibiting effect and adiponectin production promoting effect of iBright.

JP 2004-256432 A JP 2006-169181 A JP 2004-35516 A JP 2002-326947 A JP-A-9-143070 JP 2007-99651 A JP 2007-261993 A JP 2007-31302 A JP 2008-88102 A JP 2003-28111 A Japanese Patent Application Laid-Open No. 2002-284831 Nature Medicine, 2002, Vol. 8, p. 731-737 Circulation Journal, 2004, 68, p. 975-981 Nature Medicine, 2002, Vol. 8, p. 1288-1295

  An object of the present invention is to provide an effective and highly safe anti-obesity agent, lipolysis promoter, blood neutral fat elevation inhibitor, and adiponectin production promoter.

  As a result of diligent research to solve the above-mentioned problems, the present inventors have demonstrated that the extract of iBright exhibits an excellent anti-obesity effect, lipolysis promoting effect, blood neutral fat increase inhibiting effect, and adiponectin production promoting effect As a result, the present invention has been completed.

  The eyebright extract according to the present invention is obtained by extracting with a solvent from the above-ground part of eyebright. The extraction method is not particularly limited, and for example, heating extraction, room temperature extraction, or the like can be selected as appropriate.

  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, etc. Glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.) Is mentioned. These solvents may be used alone or in combination of two or more. A polar solvent such as water or a lower alcohol is preferable, and water-containing ethanol is particularly preferable.

  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.

  The above-mentioned extract may be used as it is for the anti-obesity agent, lipolysis promoter, blood neutral fat increase inhibitor and adiponectin production promoter according to the present invention, so long as the effect of the extract is not impaired. Other components can also be blended. Other components may be 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, olive oil, petrolatum, paraffin, higher alcohols and the like.

  The anti-obesity agent, lipolysis promoter, blood neutral fat elevation inhibitor and adiponectin production promoter according to the present invention can be used for any of pharmaceuticals, quasi drugs, foods and cosmetics. Examples of the administration method include oral administration (internal use) and transdermal administration (external preparation). Therefore, various preparation forms can be selected, but internal use is particularly preferable. For example, powders, granules, tablets, sugar-coated tablets, capsules, syrups, pills, suspensions, liquids, emulsions and the like. Examples of parenteral dosage forms include lotions, creams, emulsions, bath preparations, injections, and suppositories.

  The amount of the extract according to the present invention varies depending on the preparation form, purpose of use, etc., but in the case of an external preparation, the dry solid content may be 0.001 to 10% by weight, more preferably 0.01 to 5%. % By weight. In the case of internal use, the daily dose of the extract according to the present invention varies depending on the purpose of use, age, body weight, symptom, therapeutic effect, etc., but is 0.02 to 500 mg per kg body weight, particularly preferably 0. .1 to 300 mg. The addition method to the preparation may be added in advance or may be added during the production, and may be appropriately selected in consideration of workability.

  The anti-obesity agents, lipolysis promoters, blood neutral fat elevation inhibitors and adiponectin production promoters related to the present invention not only exhibit excellent effects for the purpose of each agent, but also are multifaceted with respect to obesity. Since it acts and exhibits an excellent improvement effect, it is effective in preventing and improving obesity and lifestyle-related diseases related to obesity.

  Examples are provided to describe the present invention in detail, 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.

Hot water extract of Ibright 2 kg of purified water was added to 100 g of dried Ibright (above ground), extracted at 95-100 ° C. for 2 hours, filtered, the filtrate was concentrated, freeze-dried and hot water 21 g of extract was obtained.

20% ethanol extract of iBright 1.6 kg of purified water and 0.4 kg of ethanol were added to 100 g of dried Ibright (above ground), extracted at room temperature for 3 days, filtered, and the filtrate was concentrated and frozen. Drying yielded 14 g of 20% ethanol extract.

Ethyl extract of Ibright 2 kg of ethanol was added to 100 g of dried Ibright (above ground), extracted at room temperature for 3 days, filtered, and the filtrate was concentrated and freeze-dried to obtain 9 g of ethanol extract. .

50% 1,3-butylene glycol extract of i-bright 500 g of purified water and 500 g of 1,3-butylene glycol (1,3-BG) are added to 100 g of dried i-bright (above ground) and extracted at room temperature for 7 days. And filtered to obtain 980 g of 50% 1,3-BG extract.

Lotion prescription amount (parts)
1. 50% 1,3-BG extract of iBright (Example 4) 1.0
2.1,3-BG 7.0
3. Glycerin 2.0
4). Xanthan gum 0.02
5. Citric acid 0.01
6). Sodium citrate 0.1
7). Ethanol 5.0
8). Pantothenic acid ethyl alcohol 0.1
9. Methyl paraoxybenzoate 0.1
10. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
11. Fragrance 0.1
12 Purified water 84.47
[Production Method] Components 1 to 6 and 12 and Components 7 to 11 are uniformly dissolved, mixed together, and filtered to obtain a product.

Cream prescription amount (parts)
1. Ibright ethanol extract (Example 3) 0.05
2. Squalane 5.5
3. Olive oil 3.0
4). Stearic acid 2.0
5. Beeswax 2.0
6). Octyldodecyl myristate 3.5
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Behenyl alcohol 1.5
9. Glycerol monostearate2.5
10. Fragrance 0.1
11. Methyl paraoxybenzoate 0.2
12 Ethyl paraoxybenzoate 0.05
13.1,3-BG 8.5
14 Purified water 68.1
[Manufacturing method] Components 1 to 9 are heated and dissolved and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 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.

Emulsion formulation amount (parts)
1. Ibright hot water extract (Example 1) 0.01
2. Squalane 5.0
3. Olive oil 5.0
4). Jojoba oil 5.0
5. Cetanol 1.5
6). Glycerol monostearate 2.0
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
9. Fragrance 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12 Methyl paraoxybenzoate 0.2
13. Purified water 73.19
[Manufacturing method] Components 2 to 8 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 10-13 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.

Bath preparation prescription amount (parts)
1. Eyebright ethanol extract (Example 3) 5.0
2. Sodium bicarbonate 50.0
3. Menthol 0.1
4). Yellow No. 202 (1) 0.01
5. Fragrance 0.1
6). Sodium sulfate 44.79
[Production method] Components 1 to 6 are uniformly mixed to obtain a product.

Powder formulation Formulation amount (part)
1. 20% ethanol extract of iBright (Example 2) 20
2. Dried corn starch 30
3. Microcrystalline cellulose 50
[Production method] Components 1 to 3 are mixed to form a powder.

Tablet formulation (parts)
1. Ibright hot water extract (Example 1) 1
2. Dried corn starch 29
3. Carboxymethylcellulose calcium 20
4). Microcrystalline cellulose 40
5. Polyvinylpyrrolidone 7
6). Talc 3
[Manufacturing method] Components 1 to 5 are mixed, then 10% water is added as a binder, dried after extrusion granulation. Ingredient 6 is added to the formed granule, mixed and compressed. One tablet is 0.52 g.

Tablet confection formulation amount (parts)
1. Eyebright ethanol extract (Example 3) 1.5
2. Dried corn starch 50.0
3. Erythritol 40.0
4). Citric acid 5.0
5. Sucrose fatty acid ester 3.4
6). Fragrance 0.1
[Manufacturing method] Components 1 to 4 are mixed, 10% water is added as a binder, and fluidized bed granulation is performed. Ingredients 5 and 6 are added to the formed granules, mixed and compressed. One tablet is 1.0 g.

Beverage prescription amount (parts)
1. IBRITE 20% ethanol extract (Example 2) 1.0
2. Stevia 0.05
3. Malic acid 5.0
4). Fragrance 0.1
5. Purified water 93.85
[Production Method] Components 1 to 4 are stirred and dissolved in a part of purified water of Component 5. The remaining purified water of Component 5 is then added and mixed, heated to 90 ° C. and filled into a 50 mL glass bottle.

Experimental Example 1 Confirmation of Lipolysis Promoting Effect Using Adipocytes Mouse preadipocytes (3T3-L1) were seeded in a 35 mm dish, and culture medium (Dulbecco's modified Eagle medium (DMEM medium) containing 10% fetal calf serum) ) In 2 mL, the cells were cultured at 37 ° C. in the presence of 5% CO ₂ until reaching confluence. After the cells reached confluence, the culture medium was removed by aspiration, and the cells were washed with phosphate buffered saline (PBS (−)), and then differentiation induction medium 1 (10% fetal bovine serum, 1 μM dexamethasone, 10 μg / 2 mL of DMEM medium containing mL insulin, 0.5 mM 3-isobutyl-1-methylxanthine and 100 μM ascorbic acid was added, and the cells were similarly cultured for 3 days. Next, after removing the culture medium by suction and washing the cells with PBS (−), 2 mL of differentiation induction medium 2 (DMEM medium containing 10% fetal bovine serum, 10 μg / mL insulin and 100 μM ascorbic acid) was added, Cultured for 3 days. Thereafter, the culture solution was removed by suction, the cells were washed with PBS (−), 2 mL of DMEM medium containing 10% fetal bovine serum was added, and further cultured for 8 days.
14 days after the initiation of differentiation induction, the culture medium was removed by suction, washed with PBS (−), and 2 mL of DMEM medium containing 10% fetal bovine serum was newly added. At this time, 2 μL of a sample solution obtained by dissolving 20% ethanol extract of Ibright (Nippon Powdered Drugs) in DMSO was added and further cultured for 7 days. As a control, only DMSO was added instead of the sample solution. Seven days after the addition of the sample, the medium was removed by suction and washed with PBS (−). To the dish, 1 mL of PBS (−) was added, the cells on the dish were detached, and transferred to a 1.5 mL tube. The 1.5 mL tube was centrifuged at 15,000 rpm for 5 minutes at 4 ° C., the supernatant was removed, and 0.5 mL of PBS (−) was newly added. The cell precipitate was dispersed in PBS (−) using an ultrasonic crusher, and the suspension was subjected to protein quantification and fat quantification. Protein quantification was carried out by the Raleigh method, and fat quantification was carried out using Triglyceride E-Test Wako (Wako Pure Chemical).

The amount of fat per unit protein was calculated from the results of protein quantification and fat quantification. The effect of promoting lipolysis was performed by comparing the rate of fat reduction per unit protein. That is, the higher the fat reduction rate, the stronger the effect of promoting lipolysis. The fat reduction rate was calculated by the following formula.
Fat reduction rate (%) = (1- (Amount of fat per unit protein when iBright extract is added) / (Amount of fat per unit protein when only DMSO is added)) × 100

  As a result of examining the effect of promoting lipolysis on adipocytes using the above experimental method, as shown in Table 1, the extract of eyebright according to the present invention increases the fat reduction rate in a concentration-dependent manner, and lipolysis It was found to have a promoting effect.

Experimental Example 2 Confirmation of Adiponectin Production Promoting Effect Mouse preadipocytes (3T3-L1) are seeded in a 35 mm dish, and 5% until reaching confluence in 2 mL of culture medium (DMEM medium containing 10% fetal calf serum) The cells were cultured at 37 ° C. in the presence of CO ₂ . After the cells have reached confluence, the culture medium is differentiated into differentiation-inducing medium 1 (DMEM medium containing 10% fetal bovine serum, 1 μM dexamethasone, 10 μg / mL insulin, 0.5 mM 3-isobutyl-1-methylxanthine and 100 μM ascorbic acid). And cultured for 3 days in the same manner. Thereafter, the culture medium was replaced with differentiation induction medium 2 (DMEM medium containing 10% fetal bovine serum, 10 μg / mL insulin and 100 μM ascorbic acid), and cultured in the same manner for 3 days. Further, the medium was replaced with DMEM medium containing 10% fetal bovine serum and cultured for 3 days.
Nine days after the initiation of differentiation induction, the culture medium was removed by suction, washed with PBS (−), and 2 mL of DMEM medium containing 10% fetal bovine serum was newly added. At this time, 2 μL of a sample solution obtained by dissolving 20% ethanol extract of Ibright (Nippon Powdered Drugs) in DMSO was added and further cultured for 2 days. As a control, only DMSO was added instead of the sample solution. Two days after the addition of the sample, the culture supernatant was collected and used for the measurement of adiponectin secretion. The adiponectin in the culture supernatant was quantified using a mouse / rat adiponectin ELISA kit manufactured by Otsuka Pharmaceutical Co., Ltd.

  As shown in Table 2, adiponectin production increased in a concentration-dependent manner by adding the extract of iBright related to the present invention to adipocytes. From this, it was found that the eyebright extract has an adiponectin production promoting effect.

Experimental Example 3 Confirmation of anti-obesity effect and blood lipid elevation inhibitory effect Using 20% ethanol extract of Ibright (Nippon Powdered Medicine), the anti-obesity effect and blood lipid elevation inhibitory effect were confirmed. In the experiment, 4 week old SD male rats were used as 6 rats per group. The normal food group was bred using a commercially available feed (Oriental Yeast Co., Ltd .: mouse / rat breeding-MF). As a high fat diet for inducing obesity, a high fat diet (made by Oriental Yeast Co., Ltd.) containing 23% of beef tallow was used. A high fat feed supplemented with 0.5% of 20% ethanol extract of iBright was prepared and allowed to freely feed and bred (Ibright group). The dose of the 20% ethanol extract of iBright is about 200 mg per kg body weight when calculated from the intake of feed. In addition, for comparison, a high fat diet supplemented with 0.5% casein instead of 20% ethanol extract of iBRITE (control group) and sugars that have been reported to suppress blood lipid elevation. A rearing group (sugar-transferred hesperidin group) with a high-fat diet to which 1.0% of transferred hesperidin (Hayashibara Biochemical Research Institute) was added was set.
Rats were weighed in the morning on the 12th, 16th, 23rd, 30th, 37th, 43rd and 51st days from the start of breeding. At the end of the test, dissection was performed, and histological and blood tests were performed. As an evaluation of the visceral fat accumulation amount, the weight of the epididymis fat tissue was measured. In the measurement of blood lipids, fasting was performed for 18 hours from the evening of the 47th day of breeding, and fasting blood was collected from the tail vein on the 48th day. Next, the plasma is collected by centrifugation, and triglyceride E-Test Wako (manufactured by Wako Pure Chemical Industries) is used for neutral fat, and NEFA-C-Test Wako (manufactured by Wako Pure Chemical Industries) is used for free fatty acids. Each was measured using.

  Table 3 shows the average values of the weights of each group on the 12th, 16th, 23rd, 30th, 37th, 43rd and 51st days from the start of breeding. The group to which the 20% ethanol extract of iBright was administered had a smaller weight gain than the control group, and a sufficient weight gain inhibitory effect was observed for the iBright extract. In the process of weight gain in the iBright group, no weight loss was observed in the early stages of breeding, and no histological or hematological abnormalities were observed in the anatomical findings at the end of the experiment. The iBright extract was shown to be effective in inhibiting weight gain without affecting general health.

  Table 4 shows the measurement results of blood neutral fat, and Table 5 shows the measurement results of blood free fatty acid. Breeding with a high-fat diet increased both neutral fat and free fatty acids in the blood, but it was found that any blood lipid was suppressed by administering the eyebright extract. The effect was higher and superior to glycosylated hesperidin, which has been reported to have an effect of suppressing blood lipid elevation.

  Table 6 shows the measurement results of the weight of the accessory testicular adipose tissue. Although the testicular adipose tissue weight increased by breeding with a high-fat diet, suppression of weight gain was observed by administration of an eyebright extract.

An anti-obesity agent, a lipolysis promoter, a blood neutral fat increase inhibitor and an adiponectin production promoter characterized by containing an eyebright extract related to the present invention are excellent for the purpose of each agent. Demonstrate the improvement effect. Therefore, it is useful for pharmaceuticals, quasi-drugs, foods and cosmetics for the purpose of preventing and improving obesity and lifestyle-related diseases.

Claims (5)

  An antiobesity agent comprising an eyebright extract.   A lipolysis accelerator comprising an eyebright extract. A blood neutral fat elevation inhibitor comprising an eyebright extract. An adiponectin production promoter comprising an eyebright extract. An internal preparation comprising any one of the anti-obesity agent according to claim 1, the lipolysis promoter according to claim 2, the blood neutral fat elevation inhibitor according to claim 3, and the adiponectin production promoter according to claim 4.