JP2010100530A - Anti-obestic agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an anti-obestic agent which is originated from a vegetable ingredient high in safety and is excellent in physiological actions. <P>SOLUTION: There are provided the anti-obestic agent comprising a post-heating fermented tea which is obtained by adding water in an amount of 110 pts.wt. per 100 pts.wt. of the dry weight of tea originated from Camellia sinensis and then inoculating and growing a fungus in the mixture, and has superoxide dismutase activity of ≥2,000 units/g in terms of solid content, or an extract of the post-heating fermented tea; and the anti-obestic agent, wherein obesity is visceral fat obesity or subcutaneous fat obesity. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an anti-obesity agent containing post-fermented tea having superoxide dismutase activity (hereinafter referred to as SOD activity) or an extract thereof.

Lifestyle-related diseases such as diabetes are often caused by visceral fat-type obesity in which fat has accumulated in the viscera. In addition to visceral fat-type obesity, any two or more of hyperglycemia, hypertension, and lipid abnormalities The combined state is regarded as metabolic syndrome. Many of the metabolic syndrome can cause arteriosclerosis if left untreated, leading to heart disease and stroke. Therefore, it is important to prevent metabolic syndrome.
Metabolic syndrome is said to be able to be prevented by weight loss, especially visceral fat loss, and anti-obesity agents and those having anti-obesity effects are required.

  As for anti-obesity and fat absorption suppression by tea, for example, tea drink and fat absorption inhibitor having a rebound suppression effect after dieting using puer tea extract component or jasmine tea extract component which is post-fermented tea (Patent Document 1) It has been known. In addition, a novel polyphenol compound having a lipase inhibitory activity originating in tea is provided, and a food and drink that suppresses the absorption of dietary fat and suppresses the increase of blood neutral fat is known (Patent Document 2). It has been.

  As a substance involved in the self-regulation of energy consumption, UCP (Uncoupling Protein) has attracted attention. UCP uncouples oxidative phosphorylation in the inner mitochondrial membrane and dissipates energy as heat. Have the ability to Specifically, animals that do not become obese even if they eat a lot have more UCP1 (UCP in brown adipose tissue), mice that have artificially reduced the expression of UCP1 are obese, and mice that are highly expressed can lose weight, etc. The fact is known (Non-Patent Document 1). Therefore, anti-obesity effect can be expected if UCP is highly expressed.

JP 2005-278478 A JP 2006-001909 A Masayuki Saito, "The 124th Symposium of the Japan Medical Association Symposium on Obesity", Japan Medical Association, p. 62-70, 2003

Regarding anti-obesity and fat absorption suppression by tea, the anti-obesity effect of conventional post-fermented tea represented by puer tea is not sufficient, and a plant-derived component having a stronger anti-obesity effect has been desired.
Accordingly, an object of the present invention is to provide an anti-obesity agent that is derived from a plant component with high safety and has a more excellent pharmacological action.

  The present inventors have made various studies focusing on post-fermented tea, impregnated tea with a certain amount of water, inoculated with filamentous fungi, and grown to obtain post-fermented tea with extremely high SOD activity. It was. And when the obesity suppression effect in an obesity model was investigated using the said post-fermented tea with the said high SOD activity, if this post-fermentation tea was administered, it discovered that a strong obesity suppression effect was acquired and completed this invention. . Further, it was found that if fermented tea is administered thereafter, the expression of UCP is promoted.

That is, the present invention provides a superoxide dismutase (Superoxide) obtained by adding 110 parts by weight or more of water to tea derived from Camellia sinensis, and inoculating and growing a fungus. Dismut
as) An anti-obesity agent containing post-fermented tea having an activity of 2,000 units / g or more in terms of solid content or an extract thereof. The present invention also provides the anti-obesity agent described above, wherein the obesity is visceral fat type obesity or subcutaneous fat type obesity. Furthermore, the present invention provides the above-mentioned anti-obesity agent that promotes the expression of UCP.

  The anti-obesity effect of the post-fermented tea or its extract of the present invention is strong, and obesity is particularly effective for visceral fat type obesity or subcutaneous fat type obesity. Further, the post-fermented tea or the extract thereof of the present invention promotes the expression of UCP and exhibits an anti-obesity effect. In addition, post-fermented tea or an extract thereof used in the present invention is highly safe and can be drunk or taken for a long period of time. Therefore, it is particularly useful for preventing or treating the transition to obesity.

  If the tea as a raw material of post-fermentation tea used for this invention is a Camellia sinensis seed | species, it will not be specifically limited to a kind | species etc. In addition, fresh tea leaves immediately after harvesting, tea leaves after killing blue, tea leaves after rough culm, tea leaves after twisting, tea leaves after midsummer, tea leaves after brewing, green tea such as rough tea, Semi-fermented tea and completely fermented tea can also be used for tea flowers and processed products such as these powder products, and the raw tea can be appropriately selected depending on the use of post-fermented tea. Among these, it is preferable to use non-fermented tea from the viewpoint of obtaining post-fermented tea having high SOD activity.

  The microorganism used for fermentation of these teas is preferably a filamentous fungus from the viewpoint of obtaining post-fermented tea having high SOD activity. As the filamentous fungus, it is possible to select from filamentous fungi that can be used for foods such as brewed foods and fermented foods. For example, Aspergillus genus, Eurotium genus, Rhizopus genus and the like can be used, more preferably Aspergillus oryzae, Aspergillus niger, Eurotium cristatum, Rhizopus oligosporus, and a strain that is derived from the strain that is derived from the filamentous fungus. Can also be used. These may be used alone or in combination. These strains can be obtained from strain distribution agencies such as IFO, IAM, ATCC, and NRRC, the Japan Brewing Association, and commercial seed strain sales companies.

  In the method for producing post-fermented tea used in the present invention, tea derived from Camellia sinensis is 110 parts by weight or more, preferably 150 parts by weight or more, more preferably 200 parts by weight or more, optimal for 100 parts by weight of tea dry weight. Add 300-400 parts by weight of water. The adjustment of the amount of water here means that water is added to tea used as a raw material so as to adjust to a condition that allows filamentous fungi to grow. For example, when crude tea is used, the crude tea already contains about 3 to 5% by weight of moisture, and the amount of moisture to be added needs to be adjusted in consideration of the moisture contained in the tea. Moreover, what contains 70 to 80 weight% of water | moisture content like fresh tea leaves does not need to be hydrated. The water content is important for obtaining the post-fermented tea having a high SOD activity according to the present invention, which is an amount capable of propagating or fermenting the filamentous fungi used for culture and fermentation. By preferentially propagating the filamentous fungus, contamination with various bacteria can be further prevented. It is preferable to add a sterilization step after the moisture adjustment. Examples of the sterilization conditions include heat sterilization at 80 to 100 ° C. for 30 to 60 minutes, or at 100 to 121 ° C. for 15 to 30 minutes. After the sterilization, the mixture is cooled, the tea is transferred to a sterilized culture / fermentor, and the following inoculation process is performed. The sterilization step is preferably performed in a closed system so that the moisture content of tea does not change.

  The tea whose moisture content has been adjusted is inoculated with filamentous fungi and is generally cultured and fermented at 20 to 40 ° C. for 3 to 30 days, preferably at 25 to 37 ° C. for 3 to 7 days. Here, the culture / fermentation method may be any method such as liquid culture / fermentation method or solid culture / fermentation method as long as it is a commonly used method. Then, it is preferable to add a sterilization process. The sterilization conditions include, for example, heat sterilization at 80 to 100 ° C. for 30 to 60 minutes, or 100 to 121 ° C. for 15 to 30 minutes, or heat drying at 80 to 120 ° C. Is sterilized, and the water content is 10% by weight or less, preferably 5% by weight or less. Thereby, the post-fermented tea with high SOD activity of the present invention is produced.

  The post-fermented tea of the present invention can be used as it is, but is sterilized by heating, dry heat, microwave, etc., and further pulverized, extracted in the form of powder, paste, or using a solvent, It can be used as a post-fermented tea extract. Furthermore, the post-fermented tea extract can be made into a post-fermented tea extract powder by drying and pulverizing the post-fermented tea extract using a spray dryer, drum dryer, freeze dryer, air dryer or the like. Moreover, it can be set as a granule using a granulator etc. as needed.

As the solvent used in the above extraction, water, alcohols having 1 to 6 carbon atoms or glycols can be used alone, or a combination of two or more of these solvents can be used. For example, water, ethanol, hydrous ethanol, 1, 3 -Butylene glycol, hydrous 1,3-butylene glycol, etc. are mentioned. Preferably water-containing ethanol or water-containing 1,3-butylene glycol, more preferably a mixture of water and ethanol 5 to 95: 95-5 (v / v), water and 1,3-butylene glycol 5 to 95:95 A mixture of 5 (v / v) can be used.
As extraction conditions, it is preferable to extract at about 20 to 70 ° C. for 0.1 to 5 hours, perform filtration, and cool the obtained filtrate by heating at 80 to 95 ° C. for 30 minutes.
The above extract is further purified by a known separation / purification method such as liquid-liquid separation, solid-liquid separation, filtration membrane, activated carbon, adsorption resin, ion exchange resin, etc., in order to remove inert impurities as appropriate. May be. Specifically, the extract powder may be extracted again using the extraction solvent.

  The post-fermented tea or extract thereof obtained by the above production method has an SOD activity that is one index of the antioxidant activity of 2,000 units or more, preferably 8,000 units or more, per 1 g of post-fermented tea solid content. Preferably, it has 9,000 units or more, more preferably 9,000 to 500,000 units. For example, post-fermented tea produced by the production method of the present invention using Aspergillus oryzae as a raw material tea has SOD activity of 9,000 units or more per 1 g of post-fermented tea solid content.

  The post-fermented tea having high SOD activity or an extract thereof has an excellent anti-obesity effect on an obesity model, as shown in Examples below. Specifically, these values such as body weight transition, visceral fat mass, leptin level, adiponectin level and the like, which are indicators of obesity, were significantly improved. Furthermore, the post-fermented tea or its extract having high SOD activity has been shown to promote the expression of UCP, and in particular, to promote the expression of UCP1. Therefore, the post-fermented tea or its extract is useful as a prophylactic and therapeutic agent for obesity.

  The anti-obesity agent of the present invention can be used as a pharmaceutical, food for specified health use, functional food and the like. Examples of the administration form of these drugs include oral, injection, and external use, and the oral administration form is preferred. As forms of oral medicines and foods, in addition to ampoules, capsules, pills, tablets, powders, granules, solids, liquids, gels, bubbles, etc., they can be blended in various foods. In preparing these compositions, excipients, binders, lubricants, and the like can be appropriately blended. The blending amount of the post-fermented tea or the extract thereof in these medicines and foods is preferably 0.0001 to 20% by weight, particularly 0.01 to 10% by weight, particularly 0.01% in terms of solid content (dry weight). ~ 5 wt% is preferred.

  The dosage of the anti-obesity agent of the present invention is not particularly limited, but is 10 to 5,000 mg, more preferably 50 to 1,000 mg, particularly as solid content (dry weight) of post-fermented tea or an extract per day per adult. 100 to 500 mg is preferable, and these amounts may be administered once or divided into several times.

  Hereinafter, the present invention will be described in more detail and specifically with reference to examples, but the present invention is not limited to the following examples.

[Reference Example 1]
0, 50, 70, 80, 100, 200, 300, 350, 400, and 500 parts by weight of 500 g of green tea (raw tea). And 700 parts by weight of water were added and mixed, and then heat-sterilized at 80 ° C. for 60 minutes in a closed system. After cooling to 30 ° C., the green tea leaves were transferred to a previously sterilized culture / fermentor, deposited to a thickness of 5 cm, and inoculated with Aspergillus oryzae IFO5238. After inoculation, the cells were cultured at 30 ° C. for 7 days and then dried to obtain post-fermented tea. After that, 5,000 mL of 50% (volume / volume) ethanol was added to the fermented tea, followed by extraction with stirring. Solid-liquid separation was performed by filtration, and the obtained filtrate was concentrated to a solid content of 30% (weight / volume), heated at 80 ° C. for 30 minutes and cooled to obtain a post-fermented tea extract. With respect to the post-fermented tea extract, SOD activity was measured using SOD Assay Kit-WST (manufactured by Dojin Chemical Co., Ltd.) and converted to the amount of post-fermented tea solids. The results are shown in Table 1.

The measurement of SOD activity was performed as follows. First, a post-fermented tea extract serving as a sample was diluted with distilled water, and sample solutions were prepared to be 0.075 mg / mL, 0.0375 mg / mL, 0.0075 mg / mL, and 0.00375 mg / mL. . 20 μL of this sample solution was dispensed into each well of a 96-well plate. Next, 200 μL of WST working solution prepared in accordance with Technical Information enclosed in SOD Assay Kit-WST was added and mixed well. Furthermore, 20 μL of Enzyme working solution adjusted according to Technical Information attached to SOD Assay Kit-WST was added by 20 μL each with a multi-channel pipette, incubated at 37 ° C. for 20 minutes in a plate reader, and the absorbance was measured at a wavelength of 450 nm. In addition, each was provided with a blank using the attached Dilution buffer instead of the Enzyme working solution. Further, a control using distilled water instead of the sample solution and its blank were provided. From the measured absorbance, the inhibition rate of the reaction in which “WST-1 formatzan” was produced from “superoxide” and “WST-1” was determined, and the IC ₅₀ value (unit: g / mL) of the sample was calculated.
In addition, a standard curve was prepared using a function of SOD concentration and inhibition rate, using an SOD standard product instead of the sample solution. The IC ₅₀ value of SOD (unit: units / mL) was calculated from the prepared calibration curve.
The SOD activity of the sample was calculated from the above “IC ₅₀ value of sample” and “IC ₅₀ value of SOD”. The calculation formula is as follows.
(SOD activity of sample unit: units / g) = (IC ₅₀ value of SOD) ÷ (IC ₅₀ value of sample)

  From Table 1, it can be seen that a high SOD activity is exhibited in the range of 200 to 500 parts by weight of water addition, and a higher SOD activity is exhibited in the range of 300 to 400 parts by weight.

  Moreover, when the obtained fermented tea extract was applied to the skin, there was almost no irritation. Also, the taste was good with low astringency.

  Moreover, the result of having measured the SOD activity of the post-fermented tea conventionally drunk by the same method is shown in Table 2.

  From Table 2, it can be seen that the post-fermented tea of the present invention has a higher SOD activity than the conventional post-fermented tea.

[Reference Example 2]
1,750 mL of water was added to 500 g of green tea (raw tea) and mixed, and then heat-sterilized at 80 ° C. for 60 minutes in a closed system. After cooling to 30 ° C., the green tea was transferred to a previously sterilized culture / fermentor, deposited to a thickness of 4 cm, and Aspergillus oryzae IFO5238 was inoculated. After inoculation, the cells were cultured and fermented at 30 ° C. for 7 days. After completion of the culture and fermentation, the mixture was dried with a blow dryer to obtain 486 g of post-fermented tea of the present invention. Next, 5,000 mL of water was added to the post-fermented tea, and stirring extraction was performed at 50 ° C. for 1 hour. Solid-liquid separation was performed by filtration, and the obtained filtrate was concentrated to a solid content of 30%, followed by heat treatment at 80 ° C. for 30 minutes and cooling to obtain a post-fermented tea water extract. The obtained post-fermented tea water extract was dried with a freeze dryer to obtain a post-fermented tea water extract powder (108 g). Regarding the obtained post-fermented tea water extract powder, the SOD activity was measured using SOD Assay Kit-WST. there were. The obtained extract powder was good with no astringency, no bitterness and no skin irritation.

[Example 1]
The following experiment evaluated the obesity suppression effect by the rat of the post-fermented tea water extract which is this invention.

(1) Experimental method As experimental animals, 8-week-old Wistar male rats (body weight 188-226 g / animal) were used.
Two to three animals were kept in polypropylene opaque cages (W220 × L320 × H135). The breeding room was maintained at a humidity of 40 to 50% and a room temperature of 20 to 25 ° C., and was set to a 12 hour light / dark cycle (lit; AM 8:00, extinguished; PM 8:00).

In this experiment, rats were divided into 4 groups of 6 animals, and the food and drinking water shown in Table 3 were freely fed and bred for 17 weeks. In order to cause obesity in the group (i), the other group (ii), (iii), and (iv) were mixed with 30 parts by weight of lard to 100 parts by weight of ordinary food. I gave a fat diet. The group (ii) was used as a control for high fat diet administration, and the groups (iii) and (iv) were used as evaluation groups for drinking the post-fermented tea extract in high fat diet administration. As drinking water, tap water is given to the groups (i) and (ii), and the post-fermented tea extract (obtained in Reference Example 2) is given to the group (iii) in terms of 1 kg of the rat body weight per day. Tap water containing 200 mg was given, and (iv) group was given 600 mg of post-fermented tea extract (obtained in Reference Example 2) in terms of 1 kg of rat weight per day. The contained tap water was given.
The body weight, food intake, and water consumption of each group of animals were measured daily. At the end of the administration period, the rats were sacrificed and the rats were weighed for visceral fat, examined for blood biochemistry, and examined for UCP expression.

As the above-mentioned normal food, feed MF for laboratory animals manufactured by Oriental Yeast Co., Ltd. was used. The composition was 7.7 g of water, 23.6 g of crude protein, 5.3 g of crude lipid, 6.1 g of crude ash, 2.9 g of crude fiber, and 54.4 g of soluble nitrogen-free product in 100 g.

  All results were statistically processed and expressed as mean ± standard error. The obtained data were subjected to statistical processing using one-way analysis of variance (ANOVA) followed by Turkey's multiple comparison test. For comparison between the two groups, Students t-test was used, and a risk rate of 5% or less was determined to be superior.

(2) Experimental results The body weight, food intake, and water consumption of each group of animals were measured daily, and the weekly average results are shown in FIGS. In addition, after the administration period, sacrifice was performed, and the results of measurement of visceral fat weight, blood biochemical examination, and UCP expression in the rat are shown below.

  In FIG. 1, regarding the body weight during the breeding period, the (iii) group and (iv) group administered with the post-fermented tea water extract showed a smaller value than the group (ii) which was the high fat diet administered control, and the body weight increased. It was shown to suppress. In particular, the (iv) group showed a significant difference with a risk rate of 1% at the 17th week compared to the (ii) group. The post-fermented tea water extract was excellent in suppressing weight gain and showed a remarkable obesity-inhibiting effect.

  In FIG.2 and FIG.3, there was no significant difference about the average food intake and the average water consumption of each group during the breeding period. The intake energy amount of each group and the intake amount of post-fermented tea of the present invention were as set.

  Table 4 shows the body weight ratios after 17 weeks of each group and the visceral fat mass. Body weight is expressed as mean ± standard deviation. The average body weight relative value is expressed as a relative value of each group with (ii) group being 100. The weight ratio of the visceral fat mass is expressed as a ratio to the body weight, with the visceral fat mass being the sum of the visceral fat, perirenal fat, and testicular fat.

  In Table 4, the (iii) group of post-fermented tea water extract 200 mg / kg (body weight) / day and the post-fermented tea water extract 600 mg / kg (body weight) are compared with the group (ii) which is a high fat diet administered control. ) / Day administration group (iv) all showed suppression of weight gain and visceral fat increase by high fat diet administration. In particular, the group (iv) showed a suppression of weight gain and visceral fat increase compared to the group (ii), and the weight ratio of the body weight and the visceral fat amount showed a significant difference at a risk rate of 1%. . Moreover, the body weight ratio of the visceral fat mass of the (iv) group showed a smaller numerical value than the (i) group of the normal diet administration. The post-fermented tea water extract was excellent in suppressing weight gain and increasing visceral fat, and showed a remarkable obesity-suppressing effect.

  Table 5 shows the blood biochemical test results after 17 weeks in each group.

In Table 5, about the neutral fat and blood glucose level, the (iii) group and (iv) group of post-fermented tea water extract administration showed a smaller value than the (ii) group which is a control of high fat diet administration. Regarding cholesterol, the group (iv) showed a smaller value than the group (ii).
Leptin is a hormone derived from the obesity gene, and the blood leptin concentration increases in proportion to obesity, and adiponectin is a hormone that promotes fatty acid burning and sugar uptake to improve insulin resistance.
Regarding leptin, both the group (iii) and the group (iv) showed smaller values than the group (ii). In particular, the group (iv) showed a significant difference from the group (ii). As for adiponectin, both the (iii) group and the (iv) group showed larger values than the (ii) group.
Therefore, the post-fermented tea water extract showed a remarkable obesity-suppressing effect in the blood biochemical test results.

  For UCP, the expression of UCP1, a UCP in brown adipose tissue, was examined. First, each group of individual brown adipose tissues was extracted, and then proteins were extracted from the extracted tissues, UCP1 and β-actin bands were detected by Western blotting, and these band concentrations were read as numerical values with a densitometer, The value of β-actin was shown as a relative value as a control. The larger the value, the greater the amount of UCP1.

  Table 6 shows the amount of UCP1 in the brown adipose tissue after 17 weeks of each group.

  From Table 6, regarding the expression of UCP1, (iii) group and (iv) group of post-fermented tea water extract administration are (ii) group which is a control of high fat diet administration, and (i) group of normal diet administration UCP1 increased in a dose-dependent manner and showed a UCP1 expression promoting effect. Therefore, the present invention shows a UCP1 expression promoting effect and an obesity suppressing effect.

  Table 7 shows the lipid content in the stool after drying the stool during the breeding period of each group.

  In Table 7, the fat content in the stool after drying the stool is higher in the group (ii), (iii), and (iv) of the high fat diet than in the group (i) of the normal diet showed that. Groups (iii) and (iv) of the post-fermented tea water extract administration showed that there was at least no inhibition of lipid absorption in the intestinal tract.

[Formulation Example 1]
Granules were produced according to the following formulation.
Post-fermented tea extract powder (Reference Example 2) 350 (parts by mass)
Lactose 470
Crystalline cellulose 150
Hydroxypropyl cellulose 30

[Formulation Example 2]
According to the following formulation, wet granulation was performed and tablets were obtained by tableting.
Post-fermented tea extract powder (Reference Example 2) 350 (parts by mass)
Lactose 470
Crystalline cellulose 140
Hydroxypropyl cellulose 30
Magnesium stearate 1
Talc 9

It is a figure which shows the weight change during the raising period of each group. It is a figure which shows transition of the food intake during the breeding period of each group. It is a figure which shows transition of the amount of drinking water during the breeding period of each group.

Claims (3)

  Superoxide dismutase activity obtained by adding 110 parts by weight or more of water to tea derived from Camellia sinensis seeds to 100 parts by weight of dry tea, inoculating filamentous fungi, and growing the solids. An anti-obesity agent containing post-fermented tea or an extract thereof having 2,000 units / g or more in terms of conversion.   The antiobesity agent according to claim 1, wherein the obesity is visceral fat type obesity or subcutaneous fat type obesity.   The antiobesity agent according to claim 1, which promotes the expression of UCP.

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