JP2014062052A - Insulin secretion promoter, blood glucose level elevation inhibitor and production method thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an insulin secretion promoter, a blood glucose level elevation inhibitor and a production method thereof, capable of lowering production cost and enabling use of unused royal jelly protein.SOLUTION: An insulin secretion promoter and a blood glucose level elevation inhibitor are produced by protease treatment of royal jelly protein with acidic protease, neutral protease or a combination of two or more of them. As royal jelly protein, for example, royal jelly, water-soluble royal jelly protein obtained by extracting water soluble protein from royal jelly, and alcohol denatured royal jelly protein obtained by extracting royal jelly with alcohol can be used.

Description

  The present invention relates to an insulin secretion promoter, a blood sugar level increase inhibitor, and a method for producing them.

  Prevention of illness by daily diet before getting ill is very important for a healthy life. Conventionally, there are supplements and foods containing royal jelly as a response to aging phenomena such as lifestyle-related diseases and chronic diseases.

  Royal jelly is a milky white creamy substance secreted from the mammary gland and hypopharyngeal gland of young worker bees, and is secreted as the diet of all young larvae and as the sole diet of bees that will become queen bees in the future. is there. Royal jelly has been reported to have an antiallergic action and a DNA damage inhibitory action in mice, and studies on its nutritional and pharmacological roles have also been conducted.

  Royal jelly is a substance that contains approximately 66% moisture, 14% protein, and 12% carbohydrate. Royal jelly is said to have an active ingredient of decenoic acid. When royal jelly is blended in a drink, protein is precipitated, and thus a supernatant (supernatant) from which protein has been removed is used. A part of the removed protein is marketed as a protein derived from royal jelly.

  As an existing technique, a blood glucose level increase inhibitor characterized by containing a peptide having a molecular weight of 500 to 3,000 obtained by degrading royal jelly with pepsin and pancreatin has been reported (for example, see Patent Document 1). ).

  Moreover, it is reported that the protein contained in royal jelly has an antibacterial action and an antiallergic action. Regarding the insulin action of royal jelly, decenoic acid contained in royal jelly, which is a fatty acid, has been reported as its main body (see, for example, Non-Patent Document 1 or 2).

  Hyperglycemia is a lifestyle-related disease, and if the blood sugar level is left in a high state, it becomes diabetic and blood becomes muddy. It may also cause complications such as neuropathy, retinopathy, nephropathy, cerebral infarction, angina pectoris, and diabetic gangrene. According to the 2008 survey by the Ministry of Health, Labor and Welfare, the total number of patients with diabetes is 2.37 million, and according to the “Summary of National Health and Nutrition Survey” in 2007, there are 22.1 million reserves of diabetes.

Japanese Patent No. 3691685

Okuda Takudo, Kameda Kenji, Morimoto Chie, `` Honeybee Science '', 19 (1), 9-14 (1998). Kramer KJ, Tager HS, ChildsCN, "Insulin-like hypoglycemic and immunological activities in honeybeeroyal Jelly.", 23,293-295 (1977).

  In Patent Document 1, since acidic protease pepsin and alkaline protease pancreatin are used in combination, an acidic solution and an alkaline solution are used in the production process. As a result, salt was formed, and the processing step was required, which caused a problem that the manufacturing cost was increased. Moreover, although the protein removed when the royal jelly was mix | blended with the drink was partly marketed, there existed a subject that most was not utilized. For this reason, the effective utilization method of the unused protein was sought.

  The present invention has been made paying attention to such a problem, and can reduce the manufacturing cost, and can utilize an unused royal jelly protein, an insulin secretion promoter, a blood sugar level increase inhibitor, and a production method thereof The purpose is to provide.

In order to achieve the above object, the insulin secretagogue according to the present invention is characterized by comprising a royal jelly proteolysate obtained by acidic protease, neutral protease or a combination of two or more thereof as an active ingredient.
The method for producing an insulin secretagogue according to the present invention is characterized by treating a water-soluble royal jelly protein with an acidic protease, a neutral protease, or a combination of two or more thereof. Another method for producing an insulin secretagogue according to the present invention is characterized in that an alcohol-modified royal jelly protein is treated with an acid protease, a neutral protease, or a combination of two or more thereof.

The blood sugar level elevation inhibitor according to the present invention is characterized by comprising a royal jelly protein degradation product by acidic protease, neutral protease or a combination of two or more thereof as an active ingredient.
The method for producing a blood sugar level elevation inhibitor according to the present invention is characterized in that a water-soluble royal jelly protein is treated with an acid protease, a neutral protease, or a combination of two or more thereof. Another method for producing a blood sugar level increase inhibitor according to the present invention is characterized in that an alcohol-modified royal jelly protein is treated with an acid protease, a neutral protease, or a combination of two or more thereof.

  In the insulin secretion promoter, the blood sugar level increase inhibitor and the production method thereof according to the present invention, as the royal jelly protein, for example, royal jelly, water-soluble royal jelly protein obtained by extracting a water-soluble protein from royal jelly, alcohol obtained by extracting royal jelly with alcohol Denatured royal jelly protein can be used, but is not limited thereto. A royal jelly protein degradation product is produced by treating a royal jelly protein with a protease. Proteases used in the protease treatment can be acidic proteases such as Rhizopus niveus and Aspergillus niger, neutral proteases such as Bacillus subtilis, Aspergillus oryzae, and Pig pancreas, but are not limited thereto.

  In the insulin secretion promoter, the blood glucose level increase inhibitor, and the production method thereof according to the present invention, an acidic solution and an alkaline solution are used because an acidic protease, a neutral protease, or a combination of two or more thereof is used as a protease. Unlike the case, the salt is not formed in the manufacturing process, and the salt treatment process is not required. For this reason, the manufacturing cost required for the salt treatment step can be reduced.

  The dosage form of the insulin secretagogue or blood glucose level increase inhibitor according to the present invention is preferably the most effective in the treatment, and is administered orally or parenterally such as intravenous, intraperitoneal or subcutaneous administration. Administration can be raised. Of these, intravenous administration or oral administration is preferred.

  The dosage forms to be administered include tablets, powders, granules, pills, suspensions, emulsions, soaking agents, capsules, syrups, injections, solutions, elixirs, extracts, tinctures, fluid extracts, etc. can give. Liquid preparations suitable for oral administration, such as syrups, include water, sugars such as sucrose, sorbitol, fructose, glycols such as polyethylene glycol, propylene glycol, oils such as sesame oil, olive oil, soybean oil, It can be produced using a carrier such as a preservative such as p-hydroxybenzoic acid esters, a paraoxybenzoic acid derivative such as methyl paraoxybenzoate, a preservative such as sodium benzoate, and a flavor such as strawberry flavor and peppermint.

  Also suitable for oral administration, such as tablets, powders and granules, are sugars such as lactose, sucrose, glucose, sucrose, mannitol, sorbitol, starches such as potato, wheat, corn, calcium carbonate, calcium sulfate, carbonate Inorganic substances such as sodium hydrogen and sodium chloride, plant powders such as crystalline cellulose, licorice powder, gentian powder, excipients such as paindex, starch, agar, gelatin powder, crystalline cellulose, carmellose sodium, carmellose calcium, calcium carbonate , Disintegrating agents such as sodium bicarbonate and sodium alginate, magnesium stearate, talc, hydrogenated vegetable oil, macrogol, silicone oil and other lubricants, polyvinyl alcohol, hydroxypropylcellulose, methylcellulose, ethylcellulose, potassium Merosu, gelatin, binding agents such as starch paste solution, surfactants such as fatty acid esters, can be prepared by using a plasticizer such as glycerin.

  Injectables suitable for parenteral administration preferably comprise a sterile aqueous solution containing the active compound which is isotonic with the blood of the recipient. For example, in the case of an injection, a solution for injection is prepared using a carrier comprising a salt solution, a glucose solution, or a mixture of salt water and a glucose solution. Also in these parenteral agents, the above-mentioned preservatives, preservatives, surfactants and the like can be used.

  The dose and frequency of administration of the insulin secretagogue or blood glucose level increase inhibitor according to the present invention vary depending on the administration form, patient age, body weight, nature or severity of symptoms to be treated, but royal jelly proteolysis contained therein In general, in the case of oral administration, 1 mg to 50 g, preferably 5 mg to 20 g per adult is administered once or several times a day. In the case of parenteral administration such as intravenous administration, 0.1 mg to 50 g, preferably 1 mg to 10 g per adult is administered once to several times a day. In addition, when administered to an animal, it varies depending on the age, type, nature or severity of the animal, but there is no particular limitation, and 0.1 mg to 10 g, preferably 1 mg to 1 g, per kg body weight once a day. Dosage several times. In the case of parenteral administration such as intravenous administration, 0.01 mg to 10 g, preferably 1 mg to 1 g per kg body weight is administered once to several times a day. However, the dose and the number of doses vary depending on the various conditions described above.

  The insulin secretion promoter or blood sugar level increase inhibitor according to the present invention may be added to foods and drinks or feeds, and such foods and drinks or feeds add royal jelly proteolysate to the raw materials of foods and drinks or feeds. It can be produced by a general method for producing food or drink or feed. The food / drink or feed can be shaped and granulated in the same manner as general food / drink or feed. Molding / granulation methods include fluidized bed granulation, stirring granulation, extrusion granulation, rolling granulation, airflow granulation, compression molding granulation, pulverization granulation, spray granulation, spray granulation, etc. Coating methods such as pan coating, fluidized bed coating and dry coating, puff drying, excess steam method, foam mat method, microwave heating method and other extruding methods, extrusion granulators and extruders etc. It is done.

  When the insulin secretion promoter or blood sugar level increase inhibitor according to the present invention is added to a food or drink or feed, the content of the royal jelly protein degradation product in the food or drink or feed includes: The concentration is not particularly limited as long as it has an insulin secretion promoting action or a blood glucose rise inhibiting action, but is preferably 0.001 to 100% by weight, more preferably 0.01 to 100% by weight with respect to the food or drink or feed to be contained. 0.1 to 100% by mass is more preferable.

  Specific foods and drinks to which the insulin secretion promoter or blood sugar level increase inhibitor according to the present invention is added include juices, soft drinks, soups, teas, lactic acid bacteria beverages, fermented milk, frozen desserts, butter, cheese, Dairy products such as yogurt, processed milk, skimmed milk powder, livestock meat products such as ham, sausage and hamburger, fish smelted products, egg products such as broiled and egg tofu, cookies, jelly, snacks, chewing gum and other confectionery, bread , Noodles, pickles, salmon products, dried fish, boiled salmon, seasonings and the like. Examples of the form of food and drink include powdered foods, sheet-like foods, bottled foods, canned foods, retort foods, capsule foods, tablet-like foods, liquid foods, and drinks.

  The food and drink or feed to which the insulin secretion promoter or blood sugar level increase inhibitor according to the present invention is added, if necessary, additives generally used in food or drink or feed, such as sweeteners, coloring agents, preservatives, Thickening stabilizers, antioxidants, coloring agents, bleaching agents, fungicides, gum bases, bittering agents, enzymes, brighteners, acidulants, seasonings, emulsifiers, fortifiers, manufacturing agents, fragrances, spice extracts, etc. These additives may be added.

  In the present invention, a new health food material having an action of promoting insulin secretion and an action of suppressing an increase in blood glucose level can be obtained by utilizing an unused royal jelly protein. Thereby, the royal jelly protein remaining after removing fat-soluble substances such as decenoic acid from the royal jelly can be effectively utilized. The royal jelly protein degradation product can be expected to have an insulin secretion-promoting effect, a blood glucose rise-inhibiting effect, a diabetes-improving effect, a diabetes-preventing effect, a growth-promoting effect, and an insulin-increasing effect in breast milk.

  ADVANTAGE OF THE INVENTION According to this invention, the manufacturing cost can be lowered | hung and the insulin secretion promoter, blood glucose level rise inhibitor, and those manufacturing methods which can utilize an unused royal jelly protein can be provided.

It is a graph which shows the change of the blood glucose level of a mouse | mouth after carrying out the oral gavage of the royal jelly protein degradation product of embodiment of this invention for 35 days. It is a graph which shows the change of the blood insulin level of a mouse | mouth after ingesting the royal jelly protein degradation product of embodiment of this invention freely for 35 days.

  Hereinafter, the insulin secretion promoter, the blood sugar level increase inhibitor, and the production methods thereof according to embodiments of the present invention will be described based on examples. In the following examples, representative methods for producing royal jelly protein and royal jelly protein degradation products are shown, but the present invention is not limited to this method.

[Alcohol-denatured Royal Jelly Protein and Method for Producing Alcohol-Denatured Royal Jelly Protein Degradation Product]
Royal jelly (100 g) was treated with the same amount of ethanol, and the residue was washed twice with 60% ethanol at room temperature. This residue was an alcohol-denatured royal jelly protein, which was lyophilized, and as a result, its component composition was protein (80-81%), carbohydrate (14-15%), and a small amount of lipid.

  After the obtained alcohol-modified royal jelly protein was filtered, the dried residue (12.5 g) was suspended in water (87.5 g) and adjusted to an appropriate pH. Thereafter, protease (0.25 g protease derived from Aspergillus niger) was added, and the temperature was maintained at an optimum temperature of 50 ° C., and proteolysis was performed for 5 hours. Next, enzyme inactivation was performed at a temperature of 80 ° C. for 30 minutes. The enzyme reaction solution was filtered through celite, and the filtrate was lyophilized. Thus, an alcohol-modified royal jelly protein degradation product was obtained. The yield was 8.3 g. The main component of the royal jelly protein degradation product was an enzyme degradation product (average molecular weight 879), and contained a small amount of lipid (0.9%), mineral (3.9%) and water (3%).

[Method for producing water-soluble royal jelly protein and protein degradation product of water-soluble royal jelly protein]
As a raw material other than the alcohol-denatured royal jelly protein, a method for producing a water-soluble royal jelly protein is shown below. Purified water (200 g) was added to royal jelly (50 g), and the mixture was stirred at room temperature for 30 minutes. Thereafter, centrifugation is performed at 10,000 rpm for 30 minutes, and the supernatant (first time) is recovered. Then, purified water (200 g) is further added to the precipitate, and the same operation is performed to recover the supernatant (second time). did. Each collected supernatant was freeze-dried to obtain 15.5 g of a water-soluble royal jelly protein. The yield of the water-soluble component was 87%.

  In the same manner as the method for producing an alcohol-denatured royal jelly protein degradation product from the alcohol-denatured royal jelly protein shown in Example 1, a protein degradation product of the water-soluble royal jelly protein was produced from the water-soluble royal jelly protein.

[Robust jelly protein degradation product KK-Ay oral gavage experiment]
The aqueous solution of the alcohol-denatured royal jelly proteolysate produced in Example 1 was forcibly orally administered to diabetic mice (“KK-Ay mice” manufactured by Japan SLC Co., Ltd.) for 35 days. As a control group, guava leaf extract (manufactured by Bizen Kasei Co., Ltd.) was forcibly orally administered to another diabetic mouse for 35 days. These doses are 350 mg / kg body weight and the concentration is 0.3 mL / 30 g body weight. Mice were housed in plastic cages under conditions of room temperature 25 ± 1 ° C., humidity 55% ± 5%, light / dark cycle 12 hours, and habituated for 1 week before the start of the test.

  Blood was collected every 7 days from the tail vein, avoiding fasting. The feed (“CE-2” manufactured by CLEA Japan) and water were freely consumed. The water administration group was used as a control group (control group). The blood glucose level was measured with a glucose detector and glucose test paper using mouse tail vein blood. Statistical analysis of data was performed by one-way analysis of variance and multiple comparison test (Dunnett's multiple test), with a significance level of 0.05.

  In FIG. 1, the time change of the blood glucose level of each group after completion | finish of forced administration (after 35-day progress) is shown. Each experimental result shows the average value of 5-8 mice per group. The vertical line in the graph indicates the standard error relative to the average. As shown in FIG. 1, it was confirmed that the royal jelly protein hydrolyzate administration group and the guava leaf extract administration group (positive control) significantly suppressed the blood glucose concentration (blood glucose level) compared to the control group. It was done.

[Experimental ingestion of royal jelly proteolysate KK-Ay]
The aqueous solution of the alcohol-denatured royal jelly proteolysate produced in Example 1 was mixed with a powdered feed (“CE-2” manufactured by CLEA Japan) at 0.18 w / w% (adjusted to 350 mg / kg body weight). Then, diabetic mice (“KK-Ay mice” manufactured by Nippon SLC Co., Ltd.) were allowed to freely ingest for 35 days. As a control group, guava leaf extract (manufactured by Bizen Kasei Co., Ltd.) was mixed with the powdered feed at the same ratio and allowed to freely take another diabetic mouse for 35 days. Moreover, the powder feed which does not mix anything was given as a control group (control group).

  Blood was collected every 7 days from the tail vein, avoiding fasting. The blood insulin level was measured using a mouse insulin ELISA kit. Other experimental conditions were the same as those in the gavage oral administration experiment of Example 3.

  In FIG. 2, the time change of the blood glucose level of each group after completion | finish of free intake (after 35-day progress) is shown. Each experimental result shows the average value of 6 mice per group. The vertical line in the graph indicates the standard error relative to the average. As shown in FIG. 2, it was confirmed that the royal jelly protein hydrolyzate administration group significantly increased the blood insulin concentration compared to the control group.

As shown in FIG. 2, blood insulin concentration increased when a type II diabetic mouse was allowed to freely take a royal jelly protein degradation product, and as shown in FIG. Considering that the concentration was suppressed, it can be said that the royal jelly proteolysate can be applied not only to diabetic patients whose blood insulin concentration is lowered but also as a food material for preventing diabetes.

Claims (6)

  An insulin secretagogue comprising as an active ingredient a royal jelly proteolysate obtained by acidic protease, neutral protease or a combination of two or more thereof.   A method for producing an insulin secretagogue comprising treating a water-soluble royal jelly protein with an acidic protease, a neutral protease, or a combination of two or more thereof.   A method for producing an insulin secretagogue comprising treating an alcohol-modified royal jelly protein with an acidic protease, a neutral protease, or a combination of two or more thereof.   An agent for suppressing an increase in blood glucose level, comprising a royal jelly protein degradation product obtained by acidic protease, neutral protease or a combination of two or more thereof as an active ingredient.   A method for producing a blood sugar level increase inhibitor, comprising treating a water-soluble royal jelly protein with an acidic protease, a neutral protease, or a combination of two or more thereof. A method for producing a blood sugar level increase inhibitor, comprising treating an alcohol-modified royal jelly protein with an acidic protease, a neutral protease, or a combination of two or more thereof.