WO2018101417A1

WO2018101417A1 - Gip elevation inhibitor

Info

Publication number: WO2018101417A1
Application number: PCT/JP2017/043101
Authority: WO
Inventors: 義隆古賀; 卓也森; 亮宏高村
Original assignee: 花王株式会社
Priority date: 2016-12-01
Filing date: 2017-11-30
Publication date: 2018-06-07
Also published as: JP2018090583A; JP7108397B2

Abstract

Provided is a GIP elevation inhibitor that is useful as a medicine or food and the like. The GIP elevation inhibitor contains a compressed product or extract of button mushroom as an active ingredient.

Description

GIP elevation inhibitor

The present invention relates to a GIP elevation inhibitor.

GIP (Gastrick Inhibitory Polypeptide or Glucose Dependent Insulinotropic Polypeptide) is one of the gastrointestinal hormones belonging to the glucagon / secretin family. GIP is secreted from K cells present in the small intestine by feeding lipids and carbohydrates, promotes insulin secretion in pancreatic β cells, and enhances carbohydrate and lipid uptake in adipose tissue. GIP is also known to have gastric acid secretion inhibitory action and gastric movement inhibitory action (Non-Patent Documents 1 to 3). Furthermore, it has been shown that GIP receptor-deficient mice have increased energy metabolism compared to wild-type mice (Non-patent Document 11). It has been reported that fatigue is difficult to remove due to a decrease in energy metabolism (Patent Document 5).
Therefore, suppression of the increase in GIP is considered to be effective for promoting digestion after meals, improving stomach sag, increasing energy metabolism, preventing or improving obesity, anti-fatigue, and the like.

In recent years, FABPs 4 and 5, which are isoforms of FABPs (fatty acid-binding proteins), which are known as intracellular fatty acid-binding protein families, are localized in intestinal cells that secrete GIP. Such FABP5 It has been reported that GIP secretion by lipid is reduced in non-knockout mice (Non-patent Document 4, Patent Document 1), and that a substance that inhibits FABP4 or 5 can be a GIP elevation inhibitor (Patent Document 1). .

Based on previous studies, 3-bromo-5-methyl-2-phenylpyrazolo [1,5-a] pyrimidin-7-ol (BMPP) is known as a substance that inhibits the function of GIP. Gua gum and the like are known as those that suppress secretion (Patent Document 2, Non-Patent Documents 5 to 10). In recent years, (Pro3) GIP, which is a GIP receptor antagonist, is known. However, these substances are not sufficient in terms of safety and effectiveness.

On the other hand, Tsucritake (scientific name: Agaricus bisporus) is a humus-degrading bacterium that degrades herbaceous dead grasses such as gramineous plants, and is widely used as a mushroom. Moreover, it has been reported that the mushroom extract has a leukotriene release inhibitory action and can be used as an antiallergic agent (Patent Document 3), and has an adipocyte differentiation promoting action (Patent Document 4).
However, there is no report on the relationship between the cricket mushroom pressed product or extract and GIP secretion or FABPs activity.

[Patent Document 1] JP-A-2015-194482 [Patent Document 2] International Publication No. 01/87341 [Patent Document 3] JP-A-11-217336 [Patent Document 4] JP-A-2009-263344 [Patent Document 3] [Document 5] JP 2007-308468 [Non-Patent Document 1] Brown JC et al. Canadian J Physiol Pharmacol. 1969, 47: 113-114
[Non-Patent Document 2] Falko JM et al., J Clin Endocrinol Metab. 1975, 41: 260-265
[Non-patent Document 3] Toshiji Oda et al., Gastrointestinal function and pathology, 1981, Chugai Medical Co., P205-216
[Non-Patent Document 4] Cesar A et al., Mol Endocrinol. 2014 Nov; 28 (11): 1855-65
[Non-Patent Document 5] Gagenby S J et al., Diabet Med. 1996 Apr; 13 (4): 358-64
[Non-Patent Document 6] Ellis PR et al., Br J Nutr. 1995 Oct; 74 (4): 539-56
[Non-Patent Document 7] Simoes NunesC et al., Reprod Nutr Dev. 1992; 32 (1): 11-20
[Non-Patent Document 8] Morgan LM et al., Br J Nutr. 1990 Jul; 64 (1): 103-10
[Non-patent document 9] Requejo F et al., Diabet Med. 1990 Jul; 7 (6): 515-20
[Non-Patent Document 10] Morgan et al., Br J Nutr. 1985 May; 53 (3): 467-75
[Non-Patent Document 11] Miyawaki K et al., Nat Med. 2002 Jun; 8 (7): 738-742

The present invention provides the following.
(1) A GIP increase inhibitor containing a pressed product or extract of Tsukuritake as an active ingredient.
(2) A FABP5 inhibitor containing a pressed or extracted tsukutake mushroom as an active ingredient.
(3) A food for suppressing GIP increase, which comprises a pressed or extracted tsukutake mushroom as an active ingredient.
(4) FABP5 inhibitory food comprising a crisp mushroom press or extract as an active ingredient.
(5) An energy metabolism-enhancing agent comprising a pressed or extracted tsukutake mushroom as an active ingredient.
(6) A food for promoting energy metabolism, comprising a tsukutake mushroom press or extract as an active ingredient.
(7) An obesity preventive or ameliorating agent comprising a cricket mushroom press or extract as an active ingredient.
(8) A food for preventing or improving obesity comprising a pressed or extracted tsukutake mushroom as an active ingredient.
(9) An anti-fatigue agent comprising a crisp mushroom press or extract as an active ingredient.
(10) A food for anti-fatigue comprising a pressed or extracted tsukutake mushroom as an active ingredient.
(11) Use of Tsucritake squeeze or extract to produce a GIP elevation inhibitor.
(12) Use of Tsucritake squeezed product or extract for producing FABP5 inhibitor.
(13) Use of a crisp mushroom press or extract to produce an energy metabolism enhancer.
(14) Use of a crisp mushroom press or extract for producing an agent for preventing or improving obesity.
(15) Use of a crisp mushroom pressed product or extract for producing an anti-fatigue agent.
(16) A crisp mushroom press or extract for use in suppressing GIP elevation.
(17) A cricket mushroom press or extract for use in FABP5 inhibition.
(18) A crisp mushroom press or extract for use in enhancing energy metabolism.
(19) A crisp mushroom press or extract for use in obesity prevention or improvement.
(20) A crisp mushroom press or extract for use in anti-fatigue.
(21) Use of Tsucritake squeeze or extract for suppressing GIP elevation.
(22) Use of Tsucritake squeeze or extract for FABP5 inhibition.
(23) Use of a crisp mushroom press or extract for enhancing energy metabolism.
(24) Use of a crisp mushroom press or extract for the prevention or improvement of obesity.
(25) Use of Tsucritake squeeze or extract for anti-fatigue.
(26) A method for suppressing the increase in GIP, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
(27) A method for inhibiting FABP5, comprising administering or ingesting a pressed product or extract of Tsucritake to a subject.
(28) A method for enhancing energy metabolism, comprising administering or ingesting a pressed product or extract of Tsucritake to a subject.
(29) A method for preventing or ameliorating obesity, comprising administering or ingesting a pressed product or extract of Tsucritake to a subject.
(30) An anti-fatigue method comprising administering or ingesting a pressed or extracted product of Tsucritake to a subject.

Tsukuritake squeezed product suppresses GIP increase (single dose test). A) Blood GIP concentration transition, B) Area under the curve of blood GIP concentration up to 60 minutes after administration. Tsukuritake squeezed product suppresses GIP elevation (single diet test). A) Blood GIP concentration transition, B) Area under the curve of blood GIP concentration up to 240 minutes after ingestion. Tsukuritake squeezed product suppresses GIP elevation (dose dependency test). A) Blood GIP concentration transition, B) Area under the curve of blood GIP concentration up to 120 minutes after ingestion. Tulip mushroom extract inhibits GIP increase (single diet test). Area under the curve of blood GIP concentration up to 30 minutes after ingestion.

Detailed Description of the Invention

The present invention relates to providing a GIP increase inhibitor that can be used for pharmaceuticals, foods, and the like.

The inventors of the present invention have examined materials that can control the increase in GIP, and have found that the pressed product of Tsukuritake suppresses the increase in GIP and has FABP5 inhibitory activity.

The GIP elevation inhibitor or FABP5 inhibitor of the present invention has an excellent GIP elevation inhibitory action or FABP5 inhibitory action, and is useful as a highly safe pharmaceutical or food product.

In this specification, "Tsukuritake" refers to Agaricus bisporus belonging to the genus Agaricaceae and Agaricus, and is also called "mushroom" in Japan.

As for Tsukuritake, varieties such as white, off-white, cream, brown, and Agaricus bitorquis are known, but in the present invention, the varieties are not particularly limited, and any of them is preferably used. be able to.

In the present invention, the site where Tsucritake is used is not particularly limited and may be any of an umbrella, a handle, a fruit body, a mycelium, a mycelium, or the like, but it is preferable to use a fruit body.

As the pressed product of Tsucritake, pressed juice (squeezed) obtained by pressing Tsucritake is mentioned. Although manufacture of a press thing is not specifically limited, For example, it can carry out by roughly cutting a bamboo shoot and squeezing using pressing machines, such as a slow juicer.

As an extract of Tsucritake, there is an extract obtained by extracting the fruit body of Tsucritake as it is from a crushed product, a pulverized product, or the above-mentioned pressed product.
As the solvent for extraction, either a polar solvent or a nonpolar solvent can be used. Specific examples of the solvent include, for example, water; monovalent, divalent or polyvalent alcohols; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; Cyclic ethers; Polyethers such as polyethylene glycol; Saturated or unsaturated hydrocarbons such as hexane; Aromatic hydrocarbons such as benzene and toluene; Halogenated carbonization such as dichloromethane, chloroform, dichloroethane, and carbon tetrachloride Examples include hydrogens; pyridines; dimethyl sulfoxide; acetonitrile; carbon dioxide, supercritical carbon dioxide; fats and oils, waxes, other oils; and mixtures thereof. Preferable examples include water, alcohols and aqueous solutions thereof, which include methanol, ethanol, 1,3-butylene glycol, n-propanol, isopropanol, n-butanol, isobutanol, sec-butanol, t-butanol. Etc., preferably ethanol.

The alcohol concentration (volume% at 25 ° C.) in the aqueous alcohol solution is preferably 30% by volume or more, more preferably 50% by volume or more, still more preferably 75% by volume or more, and preferably 99.5% by volume. % Or less, more preferably 99% by volume or less, and still more preferably 98% by volume or less. The concentration of the alcohol in the aqueous alcohol solution is preferably 30 to 99.5% by volume, more preferably 50 to 99% by volume, and still more preferably 75 to 98% by volume. A 30 to 99.5% by volume ethanol aqueous solution is preferable, a 50 to 99% by volume ethanol aqueous solution is more preferable, and a 75 to 98% by volume ethanol aqueous solution is still more preferable.

The amount of the solvent used in the extraction is preferably 1 to 100 mL with respect to 1 g of Tsukuritake (in terms of dry mass). The extraction conditions are not particularly limited as long as sufficient extraction can be performed. For example, the extraction time is preferably 1 hour or more, more preferably 3 hours or more, and on the other hand, preferably 2 months or less, more preferably 5 weeks or less, more preferably 2 weeks or less. The extraction temperature is preferably 0 ° C. or higher, more preferably 5 ° C. or higher, on the other hand, the solvent boiling point or lower is preferable, and 90 ° C. or lower is more preferable. Usually, extraction is performed for a long time at a low temperature and for a short time at a high temperature.
Extraction means is not particularly limited, and for example, usual means such as solid-liquid extraction, liquid-liquid extraction, immersion, decoction, leaching, reflux extraction, Soxhlet extraction, ultrasonic extraction, microwave extraction, stirring, etc. may be used. it can.

The crisp mushroom press or extract of the present invention may be a crude product as long as it conforms to, for example, a food or pharmaceutical acceptable standard and exhibits the effects of the present invention. In addition, if necessary, treatment such as removal of inert impurities, deodorization, decolorization, etc. by known techniques such as liquid-liquid distribution, solid-liquid distribution, filtration membrane, activated carbon, adsorption resin, ion exchange resin, and starch. Can be applied.
Furthermore, these purities may be increased by appropriately combining known separation and purification methods. Examples of the purification means include organic solvent precipitation, centrifugation, ultrafiltration membrane, high performance liquid chromatograph, column chromatograph and the like.

In addition, the crisp mushroom pressed product or extract of the present invention may be used as it is, may be used as a diluted solution diluted with an appropriate solvent, or may be a concentrated extract or a dry powder, or may be prepared as a paste. Good. Further, it can be freeze-dried and diluted with a solvent usually used for extraction at the time of use, for example, water, ethanol, water / ethanol mixed solution or the like. It can also be used by encapsulating in vesicles such as liposomes or microcapsules.

As shown in the examples described later, the tsukutake mushrooms have the effect of significantly suppressing the increase in GIP. In addition, the pressed product of Tsucritake has an action of inhibiting FABP5 activity.
Therefore, the pressed or extract of Tsucritake can be a GIP increase inhibitor, a FABP5 inhibitor, an energy metabolism enhancer, an obesity preventive or ameliorating agent, and an anti-fatigue agent (hereinafter also referred to as a GIP increase inhibitor, etc.) It can also be used to produce them.
Also, Tsucritake squeeze or extract can be used to inhibit FABP5, to increase energy metabolism, to prevent or ameliorate obesity, and to further prevent anti-fatigue in order to suppress GIP elevation. . Here, the “use” may be a use in a human or non-human animal, or a specimen derived therefrom, and may be a therapeutic use or a non-therapeutic use. Note that “non-therapeutic” means a concept that does not include medical practice, that is, a concept that does not include a method for operating, treating, or diagnosing a person, more specifically, a doctor or a person who has received instructions from a doctor It is a concept that does not include a method of performing surgery, treatment or diagnosis.

In the present invention, “suppression of GIP elevation” refers to suppression of elevation of GIP secreted from K cells present in the small intestine by ingesting a diet containing lipids and carbohydrates, particularly a diet rich in lipids. That is, “GIP increase suppression” in the present specification preferably means suppression of GIP increase that occurs after a meal. In addition, the “GIP increase inhibitory action” in this specification refers to a GIP secretion inhibitory action that suppresses GIP increase by suppressing GIP secretion from K cells, and suppresses GIP increase by reducing blood GIP concentration. It is a concept that includes any GIP lowering action.

In the present invention, the “GIP increase inhibitory action” can be determined based on the GIP increase inhibitory action of any oil or fatty acid. For example, the amount of GIP secretion in the test group administered or ingested with the test substance and any oil or fatty acid (for example, oleic acid), the blood in the control group to which any oil or fat or fatty acid (for example, oleic acid) was administered or ingested Compare with the amount of GIP secretion. If the test group shows a decrease in the amount of GIP secretion in the blood compared to the control group, the test substance can be evaluated as having a GIP increase inhibitory effect. In the evaluation, it is not always necessary to use a statistical method, but it is preferable to evaluate by evaluating whether there is a statistically significant difference.

Inhibition of GIP elevation suppresses the promotion of insulin secretion in pancreatic β cells and the uptake of carbohydrates and lipids in adipose tissue, and reduces the suppression of gastric acid secretion and the suppression of gastric motility (Non-Patent Documents 1 to 3). In addition, since energy metabolism is enhanced (Non-patent Document 11), the tsucritake mushroom extract or extract is used to prevent or improve obesity, promote post-meal digestion, improve stomach sag, improve gastric acid secretion, improve energy metabolism It can be used for enhancement and further anti-fatigue (Patent Document 5).

In the present invention, “FABP5 inhibition” means inhibiting FABP5 expression and inhibiting FABP5 activity, and FABP5 expression includes FABP5 gene expression or FABP5 protein expression. The FABP5 activity refers to the activity of the FABP5 protein.
The FABP5 inhibitory activity uses, for example, 1-anilinophthalene-8-sulfonic acid (1,8-ANS), which is a substrate that has binding activity to FABP5 in a hydrophobic environment and emits fluorescence when bound to FABP5. It is possible to measure by the method (refer patent document 1).

The GIP elevation inhibitor or FABP5 inhibitor of the present invention has various effects such as suppression of GIP elevation and prevention or improvement of obesity, promotion of digestion after meals, stomach sag, improvement of gastric acid secretion ability, enhancement of energy metabolism, and anti-fatigue. It can be a human or veterinary drug, quasi-drug, or food, and can be a material or preparation that is used in combination with the drug, quasi-drug, or food.
The food includes the concept of prevention or improvement of obesity, promotion of digestion after meals, stomach sag, improvement of gastric acid secretion, increased energy metabolism, and anti-fatigue. Functional label foods, foods for specified health use, foods for the sick, and supplements are included.

When the GIP elevation inhibitor of the present invention is used as a pharmaceutical (including quasi drugs), the pharmaceutical can be administered in any dosage form. Examples of the dosage form include oral administration such as tablets, capsules, granules, powders, syrups, etc. or parenteral administration such as injections, suppositories, inhalants, transdermal absorption agents, external preparations, etc. The form is oral administration.
To prepare such pharmaceutical preparations of various dosage forms, the cricket mushroom press or extract of the present invention alone or other pharmaceutically acceptable excipients, binders, extenders, disintegrations. Agents, surfactants, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, diluents and the like can be used in appropriate combinations.

When the GIP elevation inhibitor of the present invention is used as food, the form of the food is various food compositions such as breads, cakes, noodles, confectionery, jelly, frozen foods, ice creams, dairy products, and beverages. In addition to products, the same forms (tablets, capsules, syrups, etc.) as the above-mentioned oral administration preparations can be mentioned.
To prepare various forms of food, the cricket or extract of the present invention alone or other food ingredients, solvents, softeners, oils, emulsifiers, preservatives, fragrances, stabilizers, Colorants, antioxidants, humectants, thickeners, and the like can be used in appropriate combination.

The content of the crisp mushroom press or extract in the GIP elevation inhibitor of the present invention (in terms of dry matter of the extract) is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, and preferably Is 10% by mass or less, more preferably 5% by mass or less, and preferably 0.001 to 10% by mass, more preferably 0.01 to 5% by mass.

The dose or intake of the GIP elevation inhibitor of the present invention may vary according to the condition, body weight, sex, age or other factors of the subject, but if it is orally administered or ingested, the compressed product of Tsucritake per adult Or it is preferably 0.01 g or more, more preferably 0.1 g or more, and preferably 15 g or less, more preferably 6 g per day as an extract (in terms of dry matter of the extract).

Further, the GIP elevation inhibitor of the present invention is preferably administered or ingested at the time of feeding / feeding or before feeding / feeding, and in particular, administered or ingested within 5 to 30 minutes before feeding / feeding. It is preferable to do this. The subject of administration or ingestion is preferably a human having a fasting blood GIP value of 30 pg / mL or more, or a basal secretion amount of 30 mL / hour or less in a gastric juice secretion function test. In addition, humans with reduced energy metabolism, humans with obesity or their reserves who want to improve or prevent obesity, or fatigue symptoms (general malaise, slight fever, headache, lymphadenopathy, muscle pain, weakness Patients with a suspicion, disorder related to thinking ability or concentration, depressive symptoms, sleep disorders, etc., and humans who are suspected thereof are preferable.

Regarding the above-described embodiment, the following aspects are further disclosed in the present invention.
<1> A GIP increase inhibitor comprising a pressed product or extract of Tsukuritake as an active ingredient.
<2> FABP5 inhibitor containing a pressed product or extract of Tsukuritake as an active ingredient.
<3> A food for suppressing GIP increase, comprising a pressed product or extract of Tsukuritake as an active ingredient.
<4> FABP5 inhibitory food containing a pressed or extracted tsukutake mushroom as an active ingredient.
<5> An energy metabolism-enhancing agent comprising a pressed product or extract of Tsukuritake as an active ingredient.
<6> A food for promoting energy metabolism, comprising a pressed product or extract of Tsukuritake as an active ingredient.
<7> An obesity preventive or ameliorating agent comprising a crisp mushroom press or extract as an active ingredient.
<8> A food for preventing or ameliorating obesity comprising a pressed or extracted tsukutake mushroom as an active ingredient.
<9> An anti-fatigue agent comprising a crisp mushroom press or extract as an active ingredient.
<10> A food for anti-fatigue comprising a pressed or extracted tsukutake mushroom as an active ingredient.

<11> Use of a crisp mushroom press or extract to produce a GIP increase inhibitor.
<12> Use of a crisp mushroom press or extract for producing a FABP5 inhibitor.
<13> Use of a crisp mushroom press or extract for producing an energy metabolism enhancer.
<14> Use of a cricket mushroom press or extract for producing an agent for preventing or improving obesity.
<15> Use of a crisp mushroom press or extract for producing an anti-fatigue agent.

<16> A crisp mushroom press or extract for use in suppressing GIP elevation.
<17> A cricket mushroom press or extract for use in FABP5 inhibition.
<18> A cricket mushroom press or extract for use in promoting energy metabolism.
<19> A cricket mushroom press or extract for use in preventing or improving obesity.
<20> A crisp mushroom press or extract for use in anti-fatigue.

<21> Use of a crisp mushroom press or extract for suppressing GIP elevation.
<22> Use of a crisp mushroom press or extract for FABP5 inhibition.
<23> Use of a crisp mushroom press or extract for enhancing energy metabolism.
<24> Use of a crisp mushroom press or extract for obesity prevention or improvement.
<25> Use of a crisp mushroom press or extract for anti-fatigue.

<26> A method for suppressing the increase in GIP, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
<27> A method for inhibiting FABP5, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
<28> A method for enhancing energy metabolism, which comprises administering or ingesting a pressed product or extract of Tsukuritake to a subject.
<29> A method for preventing or improving obesity, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
<30> An anti-fatigue method comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.

In <31><1>,<3>,<11>,<16>,<21>,<26>, suppression of GIP increase is suppression of GIP increase due to intake of fats and oils and fatty acids.
<32> In <1> to <30>, the extract is ethanol or an aqueous ethanol extract.
<33> In <1> to <30>, the extract is a 30 to 99.5% by volume ethanol aqueous solution extract.

Hereinafter, the present invention will be described more specifically with reference to examples.
Production Example 1 Preparation of Tsucritake Pressed Product Under a 11.3 kg fruit body of Agaricus bisporus and room temperature conditions, press extraction was performed with a slow juicer (trade name: Hurom Slow Juicer HU-400). The obtained pressed product was freeze-dried to obtain 308.2 g of a pressed product powder.

Production Example 2 Preparation of Tsucritake extract 1 L of water was added to 100 g of Tsucritake pressed product, stirred at 100 ° C. for 1 hour, and extracted with hot water. The obtained extract was centrifuged (3000 rpm, 25 ° C., 5 minutes, himac CF 7D2 (Hitachi)), and the supernatant was collected and lyophilized. Of 74.7 g of the obtained dry powder, 30.0 g was suspended in 300 mL of water. To the suspension, 900 mL of 98% ethanol was added, and the mixture was allowed to stand at room temperature for 12 hours and extracted. The obtained extract was centrifuged (3000 rpm, 25 ° C., 5 minutes, himac CF 7D2 (Hitachi)), and the supernatant was collected. Ethanol in the supernatant was distilled off under reduced pressure. The remaining water was lyophilized to obtain 26.1 g of an ethanol extract.

Example 1 GIP elevation inhibitory action of Tsucritake pressed product (single administration test)
(1) Method An emulsion having the composition shown in Table 1 below was prepared using the crisp mushrooms obtained in Production Example 1 as test samples.

Eight-week-old male C57BL / 6J mice (CLEA Japan) were preliminarily raised for one week in a standard powder diet CE-2 (CLEA Japan). The breeding environment was a room temperature of 22 ± 2 ° C., a humidity of 55 ± 10%, and an illumination time of 7 o'clock to 19 o'clock. After fasting for 18 hours, groups of 6-10 animals were grouped so that their body weights were almost the same. Under isoflurane anesthesia, the test sample (Tsukuritake pressed product group, control group) was intragastrically administered to 0.02 g / g body weight. Before administration, 30 minutes and 60 minutes after administration, blood was collected from the orbit under isoflurane anesthesia (heparin-treated hematocrit micro blood collection tube, manufactured by VITREX), and blood GIP was measured. GIP was measured by ELISA (Rat / Mouse GIP (total) ELISA (Millipore)).

(2) Results FIG. 1A shows changes with time in blood GIP concentration. The area under the relative GIP blood concentration-time curve up to 60 minutes (AUC: area under the curve) is shown in FIG. 1B.
Each value is shown as mean ± standard deviation. For statistically significant differences between groups, Student's t-test for the control group was performed (*: p <0.05).

Compared with the control group, the blood GIP concentration 30 minutes after administration of the sample was significantly lower in the Tsucritake group (FIG. 1A). The area under the GIP concentration curve up to 60 minutes after sample administration was significantly lower in the Tsucritake group compared to the control group (FIG. 1B).

Example 2 GIP elevation inhibitory action of Tsucritake pressed product (single diet test)
(1) Method Mice fasted overnight (C57BL / 6J, male, 8 weeks old, n = 6-10), powdered diet (Table 2) was weighed so that calories were equal in each group, and free for 30 minutes During feeding, the high-fat diet (HF) group received 200 mg and the Tsucritake group received 252.6 mg. In addition, the Tsukuritake press thing used the thing of the manufacture example 1, and the calorie was converted into 0. Oral vein plexus blood was collected in the same manner as in Example 1 before feeding the test powder feed and at 30, 60, 120, and 240 minutes after feeding, and blood GIP was measured.

(2) Results Compared with the HF group, the blood GIP concentration at 30 minutes after feeding was significantly lower in the Tsucritake group (FIG. 2A). The area under the GIP concentration curve up to 240 minutes after ingestion of the powder feed was significantly lower in the Tsucritake group compared to the HF group (FIG. 2B).
Each value is shown as mean ± standard deviation. For statistically significant differences between groups, Student's t-test for the HF group was performed (*: p <0.05).

Example 3 GIP elevation inhibitory effect of Tsucritake press (dose-dependent test)
(1) Method Mice fasted overnight (C57BL / 6J, male, 11 weeks old, n = 6-8), powdered diet (Table 3) was weighed so that calories were equal in each group, and free for 30 minutes During feeding, the HF group received 200 mg, the 25% tsukutake added group received 250 mg, the 10% tsukutake added group received 220 mg, and the 5% tsukutake added group received 210 mg. In addition, the Tsukuritake press thing used the thing of the manufacture example 1, and the calorie was converted into 0. Oral vein plexus blood was collected in the same manner as in Example 1 before feeding the test powder feed and 30, 60 and 120 minutes after feeding, and blood GIP was measured.

(2) Results The blood GIP concentration at 30 minutes after feeding tended to be lower in the 5% tsukutake added group and the 10% tsutake mushroom added group than in the HF group, and was significantly lower in the 25% tsukutake added group. There was a dose dependency (FIG. 3A). After feeding, the area under the curve of the GIP concentration up to 120 minutes was significantly lower in the tsucritake added group than in the HF group (FIG. 3B).
Each value is shown as mean ± standard deviation. For statistically significant differences between groups, Student's t-test for the control group was performed (*: p <0.05).

Example 4 FABP5 Inhibitory Action of Tsucritake Pressed Product A pGEX-6P2 vector into which a human FABP5 gene with a Flag tag was inserted was transformed into Escherichia coli strain BL21 for expression of recombinant protein to obtain a single colony. The colony was picked up in an LB liquid medium containing 100 μg / mL ampicillin and cultured with shaking at 37 ° C. for 5 hours. Thereafter, IPTG was added so that the final concentration was 1 mM, and the cells were cultured with shaking at 37 ° C. for 3 hours, and the cells were collected. The obtained bacterial cells were solubilized by B-PER bacterial protein extraction reagent, and the soluble fraction was recovered. The E. coli soluble fraction containing the recombinant protein was subjected to GST tag cleavage and purification of the recombinant protein using GSTrap HP and PreScission protease. In a 96-well plate, the Tsucritake squeezed product (lyophilized product) prepared in Production Example 1 in a solvent (50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.5) containing the purified FABP5 protein (20 μg). It added so that it might become final concentration 0.01%. Thereafter, 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS) was added to a final concentration of 5 μM, and allowed to stand at room temperature for 15 minutes, followed by fluorescence intensity (excitation wavelength: 355 nm, Fluorescence wavelength: 480 nm).
The fluorescence intensity observed when FABP5 and 1,8-ANS coexisted was set to 100, and the loss of fluorescence when a sample was added was evaluated. The results are shown in Table 4.

A strong FABP5 inhibitory activity was observed in the Tsucritake pressed product.

Example 5
Tulip mushroom extract suppresses GIP elevation (single diet test)
(1) Method Mice fasted overnight (C57BL / 6J, male, 13 weeks old, n = 10) were weighed with powdered diet (Table 5) so that the calories were equal in each group, and allowed to eat freely for 30 minutes. In the meantime, the high-fat diet (HF) group received 200 mg, and the Tsucritake extract group received 232.5 mg. In addition, the Tsukuritake extract used the thing of the manufacture example 2, and the calorie was converted into 0. Before feeding the test powder feed and 30 after feeding, orbital venous plexus blood was collected in the same manner as in Example 1, and blood GIP was measured.

(2) Results In the Tsucritake extract group, the blood GIP concentration 30 minutes after feeding was significantly lower than that in the HF group (FIG. 4).
Each value is shown as mean ± standard deviation. For statistically significant differences between groups, Student's t-test for the HF group was performed (*: p <0.05).

Claims

GIP increase inhibitor containing tsukuritake squeezed or extract as an active ingredient.
A FABP5 inhibitor containing a pressed or extracted tsukutake mushroom as an active ingredient.
A food for suppressing GIP increase, which uses a pressed or extracted product of Tsukuritake as an active ingredient.
Food for inhibiting FABP5, which is obtained by using a pressed or extracted tsukutake mushroom as an active ingredient.
An energy metabolism-enhancing agent that uses a pressed or extract of Tsukuritake as an active ingredient.
Food for increasing energy metabolism, which is obtained by using a pressed or extracted tsukutake mushroom.
An obesity preventive or ameliorating agent comprising a crisp mushroom press or extract as an active ingredient.
Food for preventing or ameliorating obesity, which contains a pressed or extracted tsukutake mushroom as an active ingredient.
¡Anti-fatigue agent containing tsucritake mushroom press or extract as an active ingredient.
¡Anti-fatigue food containing tsucritake mushroom press or extract as an active ingredient.
使用 Use of Tsucritake squeeze or extract to produce GIP elevation inhibitor.
Use of a cricket mushroom press or extract to produce a FABP5 inhibitor.
使用 Use of a crisp mushroom press or extract to produce an energy metabolism enhancer.
使用 Use of tsukutake mushroom press or extract to produce an obesity prevention or amelioration agent.
使用 Use of tsucritake squeeze or extract to produce anti-fatigue agent.
Tsukuritake squeeze or extract for use in suppressing GIP elevation.
Tsutsutake mushroom press or extract for use in FABP5 inhibition.
Tsukuritake pressed or extracted for use in promoting energy metabolism.
Tsutsutake mushroom press or extract for use in obesity prevention or improvement.
Tsutsutake mushroom pressed or extracted for use in anti-fatigue.
使用 Use of Tsucritake squeezed or extract to suppress GIP elevation.
Use of cricket mushroom press or extract for FABP5 inhibition.
ツ Use of cricket mushroom press or extract to enhance energy metabolism.
使用 Use of cricket mushrooms pressed or extracted to prevent or improve obesity.
使用 Use of cricket mushroom press or extract for anti-fatigue.
A method for suppressing the increase in GIP, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
A method for inhibiting FABP5, comprising administering or ingesting a pressed or extracted extract of Tsucritake to a subject.
A method for enhancing energy metabolism, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
A method for preventing or ameliorating obesity, comprising administering or ingesting a pressed product or extract of Tsukuritake to a subject.
An anti-fatigue method comprising administering or ingesting a pressed or extracted product of Tsukuritake to a subject.
The agent according to claim 1, the food according to claim 3, the use according to claim 11, the compressed product according to claim 16, or the extraction, wherein the suppression of increase in GIP is suppression of an increase in GIP caused by intake of fats and oils and fatty acids. 27. The use according to claim 21, or the method according to claim 26.
The agent according to claim 1, 2, 5, 7, or 9, the food according to claim 3, 4, 6, 8, or 10, wherein the extract is ethanol or an aqueous ethanol extract, any of claims 11 to 15 The use according to claim 1, the pressed product or extract according to any one of claims 16 to 20, the use according to any one of claims 21 to 25, or the claim according to any one of claims 26 to 30. the method of.