JP7250291B2 - nutritional craving suppressant - Google Patents

Description

TECHNICAL FIELD The present invention relates to an agent for suppressing desire for nutritional supplementation.

Wheat contains starch, gluten, and the like, and the starch and gluten can be separated and recovered for use.

For example, Patent Document 1 describes an appetite suppressant containing wheat gluten or wheat gluten hydrolysate.

JP 2013-82658 A

However, in addition to Patent Document 1, further development of uses for gluten is desired.

An object of the present invention is to develop new uses for gluten separated and recovered from wheat.

The present inventors have made intensive studies to solve the above-mentioned problems, and found that peptides obtained by hydrolyzing gluten separated and recovered from wheat with a neutral protease have the effect of suppressing the desire for nutritional supplementation such as appetite. The inventors have found that the present invention has been completed.

In order to solve the above problems, the present invention includes the following aspects.
<1> A nutritional supplement desire suppressant containing, as an active ingredient, a peptide derived from wheat gluten hydrolyzate by a neutral protease.
<2> The nutritional supplement desire suppressant according to <1>, wherein the peptide is a water-soluble component of wheat gluten hydrolyzate.
<3> The nutritional supplement craving suppressant of <1>, wherein the peptide is a peptide derived from a low hydrophilic fraction among the water-soluble components of wheat gluten hydrolyzate.
<4> The agent for suppressing desire for nutritional supplementation according to any one of <1> to <3>, wherein the agent for suppressing desire for nutritional supplementation is an appetite suppressant.
<5> The nutritional supplement craving suppressant according to any one of <1> to <4>, which is for oral administration.

According to one aspect of the present invention, a novel use of gluten separated and recovered from wheat can be provided.

It is a figure which shows an example of the amino acid composition of wheat gluten. It is a figure which shows an example of the manufacturing process of a wheat gluten hydrolysis peptide. It is a figure which shows the analysis result of a wheat gluten hydrolysis peptide. FIG. 2 shows the results of SDS-PAGE of wheat gluten hydrolyzed peptides. It is a figure which shows the result of the allergen test of the wheat gluten hydrolysis peptide. FIG. 10 is a diagram showing the evaluation results of nutritional supplementation regulation of column fractions of wheat gluten hydrolyzed peptides. FIG. 2 shows the analytical results of column fractions of wheat gluten hydrolyzed peptides. FIG. 4 shows changes in the number of licks after administration of WGH-3 to mice.

[Nutrient supplement desire suppressant]
The appetite suppressant for nutritional supplementation of the present invention contains a peptide derived from wheat gluten hydrolyzate as an active ingredient. Peptides derived from wheat gluten hydrolysates are hereinafter sometimes abbreviated as "gluten-degrading peptides". Appetite etc. are mentioned as an example of a desire for nutritional supplementation.

(gluten-degrading peptide)
Gluten is a protein found in wheat and is composed of glutenin and gliadin associated with each other.

FIG. 1 shows an example of the amino acid sequence of gliadin in wheat gluten. Moreover, the ratio of glutamine (Q) contained in an example of gliadin and glutenin in wheat gluten is shown. Glutamine (Q) accounts for about 30-40% of the amino acid composition that makes up gliadin and glutenin in wheat gluten.

The gluten-degrading peptide contained in the nutritional supplement craving suppressant may be of one type, or may be a mixture of two or more types of gluten-degrading peptide.

The gluten-degrading peptide is preferably a glutamine-rich peptide in terms of enhancing the effect of suppressing the appetite for nutritional supplementation. As used herein, a glutamine-rich peptide means that the ratio of glutamine residues to the total number of amino acid residues constituting the peptide is in the range of 10% or more and 100%, preferably 30% or more and 100% or less. A certain peptide.

The molecular weight of the gluten-degrading peptide may be, for example, 300 Da or more and 15 kDa or less, preferably 300 Da or more and 10 kDa or less.

To obtain gluten-degrading peptides, commercially available gluten is hydrolyzed with a hydrolase (protease). Alternatively, gluten obtained by recovering starch from wheat may be hydrolyzed. The gluten-degrading peptide, which is the active ingredient of the nutritional supplement craving suppressant of the present invention, is hydrolyzed by a neutral protease. An example of the process for obtaining gluten-degrading peptides is shown in FIG.

Examples of neutral proteases include microorganism-derived proteases, plant-derived proteases, mammal-derived proteases, and the like. Hydrolysis conditions (eg, pH, reaction time, reaction temperature, etc.) can be appropriately selected depending on the type of neutral protease used, substrate concentration, enzyme concentration, and the like.

It is preferred to first introduce a small amount of gluten and neutral protease into near-neutral (around pH 7.0) water so as not to cause hydrolysis inhibition by dough formation. Then, it is preferable to gradually add the remaining gluten and carry out the hydrolysis treatment.

In one example, the neutral protease is used within the range of 0.1 wt% or more and 5 wt% or less based on the total weight of gluten. The amount of neutral protease used is preferably 0.2% by weight or more, more preferably 0.3% by weight or more, relative to the total weight of gluten, and the upper limit is preferably 2% by weight or less, more It is preferably 1% by weight or less or 0.7% by weight or less. The amount of the small amount of gluten added to neutral water is not particularly limited, but in one example, it is in the range of 1% or more and 10% or less of the total weight of gluten. The amount of minor gluten may preferably be 2% or more of the total weight of gluten, more preferably 3% or more, with an upper limit of 7% or less. and 5% or less is more preferable in some cases.

Gluten-degrading peptides obtained by hydrolyzing gluten preferably consist of water-soluble components. Gluten-degrading peptides composed of water-soluble components can be obtained, for example, by hydrolyzing gluten and then removing water-insoluble components by filtration or the like. It is preferable not to purify the gluten-degrading peptide with activated charcoal or the like in order to exhibit a sufficient effect of suppressing the appetite for nutritional supplementation. When purification is performed using chromatography such as reversed-phase chromatography and ion-exchange chromatography, a mixed solution of water and a lower alcohol such as methanol and ethanol may be used as the eluent. Ethanol is preferred as the lower alcohol. In addition, from the viewpoint of enhancing the effect of suppressing the appetite for nutritional supplementation, etc., the gluten-degrading peptide preferably contains a peptide derived from a low hydrophilic fraction (a peptide having low hydrophilicity) among the water-soluble components of the gluten hydrolyzate. . As used herein, the term "low hydrophilic fraction" refers to the water-soluble component of the gluten hydrolyzate containing more than 10% by mass of methanol when subjected to a Sep-Pak C18 cartridge (manufactured by Waters). It refers to the fraction eluted with a mixed solution of methanol and water.

The content of the gluten-degrading peptide in the agent for suppressing the desire for nutritional supplementation of the present invention varies depending on the form of the agent for suppressing the desire for nutritional supplementation, which will be described later. % by mass, 5 to 95% by mass, or 10 to 90% by mass.

The appetite suppressant for nutritional supplementation of the present invention is highly safe because it uses wheat, which is a cereal grain, as a raw material. In addition, as shown in Examples, hydrolysis of gluten eliminates gliadin, which is a source of allergens, and therefore is safe from the viewpoint of allergy.

(Other ingredients and dosage forms of nutritional supplement craving suppressant)
The appetite suppressant for nutritional supplementation of the present invention may further contain components other than the gluten-degrading peptide. The other ingredients are not particularly limited, but include, for example, pharmaceutically acceptable carriers, lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for adjusting osmotic pressure, buffering agents, coloring agents, flavoring agents, sweeteners, antioxidants, viscosity modifiers, and the like. In addition, if necessary, a known agent for suppressing desire for nutritional supplementation may be added as one component of the agent for suppressing desire for nutritional supplementation of the present invention to form a composite agent.

The above-mentioned pharmaceutically acceptable carrier is not particularly limited, but is a carrier that does not inhibit the effect of suppressing nutritional desire and does not exert a substantial adverse effect on the human or animal to be administered. It is preferable to have properties.

As the carrier, those already reported in this field can be widely used, and specific examples include water, various salt solutions, alcohol, vegetable oil, polyethylene glycol, gelatin, lactose, amylose, magnesium stearate, talc, and silicic acid. , paraffin, fatty acid monoglycerides, fatty acid diglycerides, hydroxymethylcellulose, polyvinylpyrrolidone, and the like, but are not particularly limited thereto. The type of carrier may be appropriately selected according to the dosage form of the nutritional supplementation suppressant, administration method, and the like.

The dosage form of the nutritional supplementation suppressant is also not particularly limited, for example, oral preparations such as tablets, pills, powders, syrups, dry syrups, liquids, suspensions, emulsions, granules and capsules; suppositories, etc. infusions; injections; and the like, preferably dosage forms for injections or oral administration. For example, oral dosage forms such as tablets and liquids are preferred from the viewpoint of portability and ease of administration.

The nutritional supplement desire suppressant of the present invention can also be used as foods such as foods with health claims such as foods for specified health uses and foods with nutrient function claims; supplements such as dietary supplements and health supplements; feeds; pet foods; .

(Administration target)
The administration subject is either a human or an animal, more specifically, any one selected from the group consisting of mammals including humans. Among them, mammals to which the nutritional supplement craving suppressant of the present invention is particularly preferably applied are not particularly limited, but experiments on mice, rats, rabbits, guinea pigs and primates other than humans, etc. animals; companion animals (pets) such as dogs and cats; domestic animals such as pigs, cows, goats, sheep and horses; humans; preferably domestic animals or humans, particularly preferably humans.

(Administration method and dose of nutritional supplement desire suppressant)
The administration method of the nutritional supplement desire suppressant of the present invention is not particularly limited, and may be systemically administered by methods such as oral administration, intravenous or intravascular administration into an artery, or enteral administration, transdermal administration, or sublingual administration. It may also be administered locally by means of administration. One preferred mode of administration is oral administration, since it is superior in terms of ease of administration and the like.

The dose of the nutritional supplementation desire suppressant may be appropriately set according to the age, sex, body weight, symptoms, administration route, administration frequency, administration period, etc. of the above-mentioned human or animal to be administered. Also, if desired, prior in vivo assays with the supplementation suppressant can be performed to determine the above dosages without undue experimentation.

The agent for suppressing appetite for nutritional supplementation of the present invention may be administered before nutritional supplementation or during nutritional supplementation.

By ingesting the agent for suppressing desire for nutritional supplementation of the present invention, it is possible to suppress the desire to supplement nutrients (nutrient components). The nutrients include carbohydrates (sugars) such as monosaccharides, disaccharides and polysaccharides; amino acids; proteins; fatty acids; lipids; By ingesting the agent for suppressing desire for nutritional supplementation of the present invention, the desire for nutritional supplementation of one or more types of nutrients can be suppressed. As shown in the examples, the agent for suppressing the desire for nutritional supplementation of the present invention is effective in suppressing the desire to supplement the combination of sugar, protein and lipid.

The nutritional craving suppressant of the present invention also functions as an appetite suppressant. When used as an appetite suppressant, it may be administered before or between meals.

EXAMPLES The embodiments of the present invention will be described in more detail below with reference to examples. Of course, the present invention is not limited to the following examples, and it goes without saying that various aspects are possible for the details. Furthermore, the present invention is not limited to the above-described embodiments, and can be modified in various ways within the scope of the claims. It is included in the technical scope of the invention. In addition, all documents described in this specification are incorporated by reference.

In the following examples, unless otherwise specified, % represents % by mass.

[Example 1] Preparation of wheat gluten hydrolyzed peptide Using wheat gluten (manufactured by Nagata Sangyo, trade name: Fumerit A2) as a substrate, according to the manufacturing process described in Fig. 2, wheat gluten hydrolyzed peptide (hereinafter referred to as , sometimes abbreviated as “WGH-2”) was prepared. As the protease, B. subtilis-derived neutral metalloprotease (manufactured by HBI, trade name: Orientase 90N) was used.

[Evaluation Example 1] Component Analysis of WGH-2 The component analysis of WGH-2 was requested to the Japan Food Research Laboratories. The analysis results are shown in FIG. As shown in FIG. 3, WGH-2 contained 77.5% protein, of which 44% was glutamine (Q) and glutamic acid (E). Due to the nature of the analytical method, glutamine and glutamic acid cannot be distinguished. However, from FIG. 1, the amino acid composition of the parent protein, wheat gluten, suggests that the substantial majority of WGH-2 is glutamine (Q).

[Evaluation Example 2] Evaluation of WGH-2 by SDS-PAGE The evaluation results of WGH-2 by SDS-PAGE are shown in FIG. No gluten was detected in the WGH-2 lane, indicating that gluten was degraded to a level undetectable by SDS-PAGE.

[Evaluation Example 3] Allergen test of WGH-2 Using a food allergen detection kit, Allergen Eye (registered trademark) Immunochromato Single Step (registered trademark) (manufactured by Primaham), WGH-2 against wheat (gliadin) An allergen test was performed. The results are shown in FIG.

Gluten was positive for wheat (gliadin), whereas WGH-2 was negative (detection limit: 2 ppm). From the results of Evaluation Example 3, it was found that WGH-2 can be labeled as gluten-free. For example, in Europe and the United States, if the gluten concentration is 20 ppm or less, it can be labeled as gluten-free.

[Evaluation Example 4] Appetite regulation effect test of column fraction of WGH-3 WGH-3 was obtained by freeze-drying the 10 kDa membrane-permeable fraction (low molecular weight fraction) of WGH-2. A 40 mg/mL WGH-3 solution was added to a Sep-Pak Vac 35 cc (10 g) C18 cartridge (manufactured by Waters) at 10 mL/cartridge and eluted with 100 mL of 0-50% MeOH aqueous solution. . The eluate was freeze-dried, and the freeze-dried product was intragastricly administered to mice using a probe at a dosage of 1 g/10 mL/kg (body weight). The number of licks of a bottle containing an aqueous solution (2% sucrose aqueous solution) placed in a cage was electrically counted and used as an index of desire for nutritional supplementation.

In addition, the lyophilized product was subjected to HPLC analysis under the following conditions. 100% eluent A was pumped for 5 minutes from the start. Then, the composition of the eluent was fed so that the composition of the eluent was changed linearly so that the eluent A became 50% eluent A/50% eluent B in 5 minutes to 25 minutes.
<Condition>
Column: TSKgel ODS-80TM (4.6 mm (ID) × 15 cm) (manufactured by Tosoh Corporation)
Eluent A: water (Milli Q water)
Eluent B: MeOH
Flow rate: 1.2 mL/min Temperature: 40°C

The evaluation results are shown in FIG. In FIG. 6, for example, "WGH 0-10%" indicates the results of 0-10% MeOH elution fractions. As shown in Fig. 6, in the fractionation with the Sep-Pak C18 cartridge, it was found that the 0-10% MeOH elution fraction (highly hydrophilic fraction) increased the number of licks and tended to promote nutritional supplementation. rice field. The 0%-10% MeOH elution fraction eluted from 0-15 minutes in HPLC analysis. On the other hand, in the MeOH elution fraction (low hydrophilic fraction) of more than 10% to 50%, it was found that the number of licks decreased and there was a tendency to suppress the desire for nutritional supplementation. The 10-50% MeOH elution fraction eluted at 15-35 minutes in HPLC analysis.

[Evaluation Example 5] Analysis of column fractions of WGH-2 A 20 mg/mL WGH-2 solution was continuously applied to Diaion HP20 (manufactured by SIGMA-ALDRICH, bed volume: 125 mL). Then, 50 mL of the eluate was collected. The results of HPLC analysis of the recovered eluate are shown in FIG. The HPLC analysis conditions are the same as in Evaluation Example 4. For WGH-2, the same evaluation test as in Evaluation Example 4 was performed, and the number of times of licking was measured to evaluate the nutritional supplementation tendency. The promoting tendency and suppressing tendency described in FIG. 7 are the results of evaluating the nutritional supplementation tendency for WGH-2.

As shown in FIG. 7, in the HPLC analysis of the WGH-2 solution, the fraction eluted at 15 to 35 minutes (more than 25% to 50% MeOH elution fraction) showed a tendency to suppress nutritional supplementation. Do you get it. On the other hand, the HPLC analysis of the eluate collected by continuously subjecting the WGH-2 solution to Diaion HP20 did not show any peaks corresponding to this component that tends to suppress the appetite for nutritional supplementation.

[Evaluation Example 6] Investigation of Presence or Absence of Appetite Suppressing Effect WGH-3, which is a 10 kDa membrane-permeable fraction (low molecular weight fraction) of WGH-2, was examined for an appetite suppressing effect. Eight male 5-week-old C57BL6 mice were used to electrically measure the number of times they licked a bottle containing an aqueous solution (aqueous solution containing 2% sucrose, 2% albumin and 1% intralipos) placed in a cage. as an indicator of Test samples (WGH-3 or physiological saline) were administered intragastrically via a probe at a dosage of 3 g/10 mL/kg body weight. Physiological saline was used as a control. For 10 minutes from 30 minutes after administration to 40 minutes after administration, the cumulative number of licks was measured over time using a lick measurement device (manufactured by Melquest). The evaluation results are shown in FIG.

As shown in FIG. 8, as a result of measuring the cumulative number of licks, the effect of suppressing appetite (desire for nutritional supplementation) was observed when WGH-3 was administered compared to when physiological saline was used as a control. On the other hand, when a 2% sucrose aqueous solution was used in place of the 2% sucrose, 2% albumin and 1% intrapolis-containing aqueous solutions, it was confirmed that the 2% sucrose solution promotes the desire for nutritional supplementation.

INDUSTRIAL APPLICABILITY The present invention can be used as an appetite suppressant or an appetite suppressant for nutritional supplementation.

Claims (3)

Contains a neutral protease-treated wheat gluten as an active ingredient,
The neutral protease-treated product contains a peptide,
The peptide, among the water-soluble components of wheat gluten hydrolyzate, when subjected to Sep-Pak C18 cartridge (manufactured by Waters), peptides derived from fractions extracted by more than 10% to 50% aqueous methanol solution is a nutritional craving suppressant. 2. The nutritional supplementation desire suppressant according to claim 1 , wherein the nutritional supplementation suppression agent is an appetite suppressant. 3. The nutritional supplement craving suppressant according to claim 1 or 2 , which is for oral administration.

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