TW201127391A - Cholecystokinin secretion promoter - Google Patents

Abstract

According to the present invention, a cholecystokinin secretion promoter is provided, which comprises potato extracts. The cholecystokinin secretion promoter of the present invention can be used as a mean for providing satiety and appetite inhibition, and is suitable to be used as pharmaceutical and refresher.

Description

201127391 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to a cholecystokinin secretion promoting agent. [Prior Art] In recent years, people who are worried about metabolic syndrome have gradually increased due to environmental changes at various levels. As metabolic syndrome increases the risk of various diseases, its improvement is a social issue. As a preventive method, there are examples of improvements in eating and drinking, moderately sustained exercise, and control of calorie intake. However, the improvement of eating and drinking life requires special knowledge, and it is not easy to obtain sufficient time to exercise moderately. Therefore, the control of calorie intake is the easiest and most convenient way to lose weight. To control calories, a method of reducing the amount of meal per meal can be cited. However, reluctance to reduce the amount of meals is accompanied by pressure and may also be the cause of the rebound. In order to reduce the amount of meals without feeling stressed, even a small amount of meals can be satiety, and it is important to use the feeling of fullness after a meal. Due to the ingestion of food, various peptides are stimulated in the body to stimulate satiety. Cholecystokinin (hereinafter also referred to as "CCK") is an important controlling factor for satiety in humans. The release of CCK after feeding is caused by delaying the emptying of gastric contents and activation of receptors in the appetite-dependent brain. Various satiety effects. Therefore, it is possible to maintain a relatively high concentration of CCK in the blood under physiological conditions, and it is possible to make a relatively small amount of dietary intake by satiety, and it is expected to play an important role in the prevention of obesity. However, CCK itself has the disadvantage of being activated by the enzyme of the stomach. 201127391 Disadvantages, in order to directly increase the concentration of CCK, it has to rely on blood. Therefore, there is an expectation that the development of a means for promoting secretion of CCK by a certain foraging factor by a certain foraging factor is expected. As a dietary factor that promotes CCK secretion, various ingredients or constituent properties have been reported. For example, Patent Document 1 describes a composition containing a) a protein selected from the group consisting of casein, whey, and soybean; b) a glycomacropeptide or casein macropeptide; c) a long-chain fatty acid. (Ci2 to C18); and d) soluble fiber or insoluble fiber or a mixture thereof; Patent Document 2 describes a pepsin decomposition product of soybean/S-conglycinin; and Patent Document 3 describes a milk A protein and a whey protein hydrolyzate are disclosed; in Patent Document 4, a whey protein hydrolyzate is described; and in Patent Document 5, a use of a peptide derived from a milk protein hydrolysate is described. Patent Document 6 discloses an oral administration of a trypsin inhibitor which stimulates the release of CCK to enhance satiety. Trypsin inhibitors are presumably acted by a negative feedback signal that inhibits CCK secretion. According to this method, the trypsin inhibitor maintains the CCK concentration and thereby maintains a feeling of satiety. Patent Document 7 discloses a potato protease inhibitor II (PI2) which increases the concentration of CCK in plasma. Patent Document 8 describes an extract or extract obtained by extracting hot water from yeast, which promotes CCK secretion. PRIOR ART DOCUMENT Patent Literature Patent Document 1: Japanese Patent Publication No. 2003-523368

S -4- 201127391 Patent Document Patent Document Patent Document 4 Patent Document 曰本特开2004- 10569 曰本本表 2005_538704号曰本本表 2006-510367 曰本本表 2006-514083号 Patent Literature 6: U.S. Patent No. 4,491,578, the entire disclosure of which is hereby incorporated by reference. Problem The object of the present invention is to provide a factor useful for promoting secretion of cholecystokinin, which is an important controlling factor for satiety. Means for Solving the Problems The present invention provides a cholecystokinin secretion promoting agent which contains a potato extract. EFFECTS OF THE INVENTION According to the present invention, a novel cholecystokinin secretion promoting agent is provided. The cholecystokinin secretion promoting agent can be used as a means for giving a feeling of satiety, inhibiting the stomach of I#, and is suitable for use in medicines and foods and drinks. [Embodiment] Embodiments of the invention Hereinafter, embodiments of the invention will be described. Further, the explanation of the stomach sputum should not be limited to the following embodiments, and various changes can be made within the range described in the patent stomata f ^ s.

S 201127391 There is no particular limitation on the potato variety that is the raw material of potato extract. For example, Baron Potato, May Queen, Kita (Kitaakari; Nonglin 29), Toya (Touya; Nonglin No. 31), Feng White (1'〇7.〇51^〇;Nonglin No.21), Inka's Awakening (Nonglin No.44), Out Island, Tensheng Xiaojin (Nonglin No.41). The origin of the potato is also not particularly limited, but the potato produced in Hokkaido is preferred. In addition, it is better to use raw potatoes (for example, unprocessed within 2 inches) as raw materials. The method for producing the potato extract is not particularly limited, and an extraction method, an extraction solvent, a production aid, or the like which can be used in the production of general foods can be used. - In the embodiment, the potato extract can be modified as described below. First, an extraction solvent (acid) is added to a liquid material obtained by pulverizing and extracting an edible portion (tuber) of potato, and the pH is adjusted to be acidic. Second, solvent extraction can be carried out under heating. It can be used as a solvent for extraction, and generally, hydrochloric acid, acetic acid, sulfuric acid, formic acid, citric acid, ascorbic acid (vitamin C) and the like can be mentioned. The amount or concentration of the acid to be added can be appropriately set so that the pH is generally 2 to 5, and preferably 3 to 4 or more. The solvent extraction is carried out, generally 10 to 60 minutes at 70 to 90 ° C, and preferably 10 to 20 at 75 to 85 ° C. After the solvent extraction, the precipitate or the insoluble fraction may be removed by a suitable separation means such as centrifugation or filtration to recover the supernatant or the soluble fraction. Further, the same extraction treatment may be performed again on the precipitate or the insoluble portion after the extraction, and the supernatant or the soluble fraction may be recovered. 1 201127391 Self-recovering supernatant or soluble fraction (ie potato extract), 'Separate and recover more than 10,000 molecular weight by membrane treatment. Next, sodium hydroxide is added to the above-mentioned extract after cooling, and the pH is adjusted to be neutral or in the vicinity thereof to obtain a horsebell extract. The membrane treatment for molecular weight fractionation may be an ultrafiltration membrane treatment or a gel filtration membrane treatment, and it is preferred to treat it with an ultrafiltration membrane. The potato extract is further available for treatment by means of drying, pulverization, and the like. As the drying method, any conventional method can be used, and examples thereof include air drying, heat drying, spray drying, and freeze drying. The potato extract may be added with, for example, an excipient (e.g., dextrin), and dried by a spray drying method or the like. The pulverization method includes, for example, means of micro pulverizing or micronizing using a pulverizer or an attritor. The potato extract may be either in the form of a liquid or a solid after removal of the solvent. The present invention is preferably a potato extract prepared by Toyo New Drug Co., Ltd. Potato extract can directly stimulate and promote the secretion of CCK. Regarding the promotion of CCK secretion in animals (including humans), the inhibition of CCK secretion inhibition by conventional methods is by the action of trypsin which inhibits the CCK secretion inhibitor. However, according to the potato extract, it is expected to have a stimulation and promotion effect on the production and secretion of CCK from cells which can produce CCK. The CCK secretion-promoting effect is attributed to the composition of the potato extract, 201127391. It is adsorbed to the adsorbent HP20 and dissolved in 80% (v/v) ethanol. According to the present invention, the potato extract can be used as a CCK secretion promoting agent, and a CCK secretion promoting agent containing a potato extract is provided. The CCK secretion-promoting agent of the present invention, by administration or ingestion, can give a subject (including a human) a feeling of satiety and suppress appetite. The CCK secretion promoting agent of the present invention is also useful for the prevention and treatment of any symptom which is improved by an increase in CCK secretion. The CCK secretion promoting agent of the invention may be used alone or in combination with various nutrients, excipients, extenders, adhesives, tackifiers, emulsifiers, coloring materials, spices, food additives, nutritional supplements, seasonings. Etc., to prepare a composition for oral administration or ingestion. The compounding amount of the CCK secretion promoting agent of the present invention may be determined according to the type or dosage form of the product to be mixed; the age, sex, body weight or state of the subject to be administered or ingested; the method, period or time of administration or ingestion, etc. set up. When the administration amount of the CCK secretion promoting agent of the present invention is, for example, a food intake such as a health food, the active ingredient is usually 10 to 200 mg per person per day (times per person per person), and is 100. ~ lOOOOg is preferred, especially 3 00~lOOOOg. The CCK secretion promoter may be administered before meals, between meals, and after meals, preferably within 2 hours before meals, shortly before meals, or immediately after meals. It can also be divided into several rounds. The composition for oral administration or ingestion can be formed into a dosage form such as a hard capsule, a capsule of a soft capsule, a lozenge, a lozenge, etc., in accordance with the preference of the person in need, 201127391 or powdery, granular, scorpion, etc. The shape. It can also be formulated as a liquid dosage form such as a solution, suspension, or emulsion. The CCK secretion promoting agent of the present invention can be used as a medicine, a nonmedicinal product, a food for specified health uses, a nutritional supplement, other foods, or the like, or Mix and use here. The composition for oral administration or ingestion, or a mixed product thereof, is preferably directly consumed according to the dosage form or shape or preference, and is preferably dissolved in water, hot water, milk, soy milk, tea, juice, etc. Hereinafter, the present invention will be specifically described based on examples, but the present invention is not limited to the examples. EXAMPLES In the following examples, potato extracts made from Toyo New Drug Co., Ltd. were used as potato extracts. (Example 1: Appetite suppression of potato extracts.) For potato extract and soybean trypsin inhibitor (S BTI : Sigma Type II-S; Sigma), the appetite suppression of rats was investigated. effect. It has been reported that SBTI increases the release of CCK, and as a result, the food intake is lowered (Patent Document 6). The order of the appetite suppression test is as follows: Animal: SD strain male rats, 8 weeks old feed: AIN-93G (according to Reeves PG, Nielsen FH, Fahey GC Jr. AIN-93 purified diets for laboratory rodents: final report 201127391 of the American Institute 〇f Nutrition ad hoc wi committee on the reformulation of the AIN-76A rodent d Nutr 1 23: 1 939-5 1,1 993 modulation) shading cycle (reverse conversion): dark period 10: 00-22: 00 Ming 22 : 00-10 : 00 1 2 hours after fasting, shortly before entering the dark phase, the test substance solution of 2 m 1 of dissolved water was administered to the rats by mouth (intragastric) with a feeding tube. After that, silver food AIN - 9 3 G feed. After 1, 2, 3, and 6 hours after hungry feed, feed straight. As a test substance, each system of the rat used 100 mg of potato extract or S Β ΤΙ (only water was administered as a negative control group). The result is shown in Fig. 1. Fig. 1 is a bar graph showing the food intake of rats after 1, 2, 3 and after feeding of the feed treated with potato extract and soybean trypsin inhibitor. The food intake is expressed in g on the vertical axis, and the elapsed time (hours) after feeding the feed is shown on the horizontal axis. The results from the left control group in the middle group indicate the negative control group (water only), SBTI treatment, and potato extract treatment. The knot 値 is the mean 値 + standard error (n = ll). In the figure, a and b are (Duncan)'s multivariate domain significant difference analysis, which is represented by the symbol with the phase of the mother, to show a significant difference in the treatment group within the same elapsed time group (P < 0.05). As shown in Fig. 1, compared with the negative control group in which only water was administered, the administration significantly reduced the food intake after 1 to 6 hours. The horse extract also caused a decrease in food intake, and after 1 hour and 3 small iting iet. J 丨 period: solution, and then measured 6 small units of horse bell treatment shown in the KENTI bell between the Duncan characters After the potato -10- 201127391 visible significant reduction in food intake. Further, the protein content, relative to SB ΤΙ, was a purified product of the test grade, and the potato extract was 20% (determined according to the Kjeldahl method). As described above, a continuous decrease in food intake was observed when both the potato extract and the SBTI were administered at 100 mg. (Example 2: CCK secretion promoting action of potato extract) The CCK secretion test was carried out according to the following procedure. First, CCK-producing cell line STC-1 from mouse small intestine (by Dr, D. Hana'han, University of Califoni) , supplied by San Francisco, CA), in a 48-well dish, in DMEM (Dulbecco's Modified Eagle Medium) medium containing 10% fetal bovine serum (FBS), in the presence of 37 ° C, 5% CO 2 , culture 2~ On the 3rd, it became a subconfluent cell. Next, the medium was removed from the wells and Hepes buffer (140 mM sodium chloride, 4.5 mM potassium chloride, 20 mM Hepes, 1.2 mM calcium chloride, 1.2 mM magnesium chloride, 10 mM D-glucose, 0.1% bovine serum albumin) in the wells. (BSA), pH 7.4) After washing the cells, a test substance solution 100#1 dissolved in Hepes buffer was added to each well, and cultured at 37 ° C for 60 minutes. The supernatant was recovered, and the cells were pelleted by centrifugation at 800 x g for 5 minutes at 4 ° C, and the supernatant 80 # 1 was recovered and stored frozen. The supernatant after cryopreservation was appropriately thawed, and the CCK concentration in the supernatant was quantified using a commercially available enzyme-linked immunosorbent assay (ELISA) kit (manufactured by Phoenix Pharmaceuticals Inc.). The CCK concentration (pM) in the above supernatant was taken as the amount of CCK secreted from the CCK-producing cell line STC-1.

S -11- .201127391 The following were used as test substances: potato extract (1, 5, 10 and 20 mg/ml); SBTI (Sigma Type II-S; manufactured by Sigma; 1, 5, 10 and 20 mg/ml) And soybean/5 conglycinin peptone (BconP: soy/3 conglycinin pepsin hydrolysate: mix /3 conglycinin (manufactured by Fuji Oil Co., Ltd.) with phosphoric acid solution And adjusted to pH 1.85, and added with 0.5% by mass of pepsin (Sigma), and reacted at 37 ° C for 10 minutes, after inactivation of pepsin by boiling, the supernatant was centrifuged. Neutralize and remove salt; 5mg/ml; used as a positive control group). The result is shown in Fig. 2. Figure 2 shows the CCK secretion of CCK-producing cell line STC-1 from mouse small intestine treated with potato extract, soybean trypsin inhibitor, and soybean/5-glycinin, respectively, as a histogram. Picture. The CCK secretion was expressed in the pM unit on the vertical axis, while on the horizontal axis, the "CN" system was the result of the negative control group (no test substance added); the "BconP" was the 5 mg/ml soybean yS conglycinin gland treatment group. Results: "S BTI" was the result of the soybean trypsin inhibitor treatment group (1 '5, 10, and 20 mg/ml), and the "Maling summer stroke" was the Maling summer stroke treatment group (1 , 5, 10 and 20 mg/ml) results. The number of results 値 is the mean 标准 + standard error (n = 3 to 4). In the figure, a, b, and c are Duncan's multivariate domain significant difference analysis, which are represented by symbols with distinct letters to show significant differences between treatment groups (P < 0.05). -12-i 201127391 As shown in Fig. 2, the potato extract showed a concentration-dependent CCK secretion-promoting activity relative to the CCK secretion-promoting activity of SBTI. The potato extract showed the same level of CCK secretion-promoting activity as the positive control group BconP at a concentration of 5 mg/ml. It was found that the release of CCK released from the reported trypsin inhibitor SBTI did not show CCK secretion-promoting activity. Potato extract has strong CCK secretion promoting activity. Further, the trypsin inhibitory activity was investigated for the potato extract and S BTI using the synthetic substrate benzidine arginine-p-nitroanilide (ΒΑΡΝΑ). Among the 50% inhibition (IC50), SBTI has a trypsin inhibitory activity of 10 times or more (20 to 30 times) of potato extract. Even SBTI 50 mg has a trypsin inhibitory activity higher than that of potato extract of 100 mg. Therefore, the CCK secretion promoting effect of potato extract may contribute to factors different from trypsin inhibitory activity. (Example 3: CCK secretion promoting action of the potato extract fraction according to the column chromatography using the adsorbent HP20) The same test as in Example 2 was carried out except that the following were used as the test substances. The test materials used in this example were as follows: potato extract (5 mg/ml); potato extract HP20 non-adsorbed fraction (3.72 mg/ml; equivalent potato extract 5 mg/ml portion); potato extract HP20 adsorption-20 % ethanol dissolution fraction

S -13- 201127391 (0.5 9 mg / m 1 ; equivalent potato extract 5 mg / m 1 part): and potato extract HP20 adsorption - 80% ethanol elution fraction (0.23 mg / ml; equivalent potato extract 5 mg / Ml)). The modulation of the above test substances is explained below. The potato extract was passed through, adsorbed on a column packed with Diaion HP20 (manufactured by Mitsubishi Chemical Corporation), and washed with pure water. Secondly, 20% (v/v) ethanol and 80% (v/v) ethanol were sequentially passed through the column to dissolve the adsorbate. Combining the non-adsorbed liquid flowing out of the column adsorption operation with the washing liquid discharged by pure water washing; by 20% (v/v) ethanol eluent; and by 80 The % ( v / v ) ethanol solution is concentrated and lyophilized to make the HP20 non-adsorbed part of the potato extract, the potato extract HP20 adsorbs -20% ethanol, and the potato extract HP20 adsorbs -80% Ethanol elution fraction. The recovery rates correspond to 74.4%, 11.7% and 4.5%, respectively, of the potato extract used for grading. This result is shown in Figure 3. Figure 3 will be treated with potato extract, potato extract HP20 non-adsorbed fraction, potato extract HP20 adsorbed -20% ethanol elution fraction, and potato extract HP20 adsorption-80% ethanol elution fraction. The CCK secretion amount of the CCK-producing cell line STC-1 of the mouse small intestine is shown in a histogram. The amount of CCK secretion is expressed in the pM unit on the vertical axis, and on the horizontal axis, "Blank" indicates the result of the negative control group (no test substance added); "Potato extract" is the result of the potato extract treatment group; "HP20 "non-adsorbed part" is the result of the potato extract HP20 non-adsorbed part treatment group; "HP20 adsorption -20%

S -14- 201127391 Ethanol Separation Part" is the result of the potato extract HP20 adsorption--20% ethanol elution fraction treatment group; and "HP20 adsorption-80% ethanol dissolution fraction" is the potato extract HP20 adsorption-80% ethanol The results of the fractionation treatment group. The number of results is 値 + standard error (n = 3 to 4). In the figure, a, b and c are Duncan's multivariate domain significant difference analysis, which are represented by symbols with distinct letters to show significant differences between treatment groups (P < 0.05). The potato extract HP20 adsorption-80% ethanol fractionation treatment group, although only a low concentration of 0.23 mg/ml, was observed to promote high CCK secretion promoting activity. Potato extract HP20 non-adsorbed fraction treatment group and potato extract HP20 adsorption--20% ethanol fractionation treatment group showed no significant CCK secretion promoting effect. (Example 4: Concentration dependence of CCK secretion promoting action on potato extract HP20 adsorption-80% ethanol elution fraction) The same test as in Example 2 was carried out except that the following was used as the test substance. The test materials used in this example were as follows: Potato extract (5 mg/ml); Potato extract HP20 adsorption - 80% ethanol elution fraction 〇 225 mg/ml (modulated in the same manner as in Example 3; equivalent potato extract 5 mg) /ml portion) 0.113mg/ml (The above 0.225mg/ml preparation is diluted 2 times in Hepes buffer)

S -15- 201127391 This result is shown in Figure 4. Figure 4 is a CCK-producing cell line from the mouse small intestine treated by potato extract (5 mg/ml), and 0.225 mg/ml or 0.113 mg/ml potato extract HP20 adsorption-80% ethanol. The amount of CCK secreted by STC-1 is shown in a histogram. The amount of CCK secretion is expressed in PM units on the vertical axis, and on the horizontal axis, "Blank" indicates the result of the negative control group (no test substance added); "Potato extract 5 mg/ml" is potato extract (5 mg/ml) The results of the treatment group; "0.113 mg/ml" of "HP20 adsorption-80% ethanol elution fraction" is the result of the 0.113 mg/ml potato extract HP20 adsorption-80% ethanol elution fraction treatment group; and "0.225 mg" /ml" is the result of the 0.225 mg/ml potato extract HP20 adsorption-80% ethanol elution fraction treatment group. The number of results 値 is the mean 标准 + standard error (n = 3 ~ 4). In the figure, a, b and c are Duncan's multivariate domain significant difference analysis, which are represented by symbols with distinct letters to show significant differences between treatment groups (P < 0.05). The potato extract HP20 adsorption-80% ethanol elution fraction of CCK secretion promotes concentration-dependent. The potato extract HP20 adsorbed -80% ethanol, and even at a low concentration of 0.11 3 mg/ml, CCK secretion-promoting activity comparable to that of the potato extract (5 mg/ml) treatment group was observed. Industrial Applicability The potato extract can be used as a means of suppressing appetite by giving a target animal (including humans) which is administered or ingested with potato extract to a fullness according to its secretion promoting effect of cholecystokinin. . The cholecystokinin secretion promoter containing such a potato extract can be widely used in medicines and diets -16 - i 201127391. Moreover, the insights of the present invention may also be useful for improving metabolic syndrome that is also a social issue. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a bar graph showing the food intake after 1, 2, 3, and 6 hours after feeding of potato extract-treated rats and soybean trypsin inhibitor-treated rats. Figure 2 is a bar graph showing the amount of CCK secreted by the CCK-producing cell line STC-1 from the mouse small intestine treated with potato extract, soybean trypsin inhibitor, and soybean/3 conglycinin gland, respectively. . Figure 3 shows the treatment by potato extract, potato extract ΗP20 non-adsorbed fraction, potato extract HP20 adsorbed -20% ethanol elution fraction, and potato extract ΗΡ20 adsorption-80% ethanol. A histogram of the amount of CCK secreted by the CCK-producing cell line STC-1 of the mouse small intestine. Figure 4 shows CCK production from mouse small intestine treated with potato extract (5 mg/ml) and 0.225 mg/ml or 0.113 mg/ml potato extract HP20 adsorption-80% ethanol elution fraction. A histogram of the amount of CCK secreted by the cell line STC-1. [Main component symbol description] None.

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Claims (1)

201127391 VII. Scope of application: 1. A secretion promoter for cholecystokinin, which contains potato extract. S -18-