JP2005200386A - Medicine having lipase blocking effect, fat absorption inhibitory effect and cholesterol absorption inhibitory effect - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a medicine that has excellent lipase blocking effect, fat absorption inhibitory effect and cholesterol absorption inhibitory effect with reduced harsh taste, thus can be easily added to foods and being processed, further has bile acid reabsorption inhibitory effect and can easily be ingested with high safety, and provide foods each having excellent lipase blocking effect, fat absorption inhibitory effect and cholesterol absorption inhibitory effect by containing the medicine. <P>SOLUTION: The medicine contains a polysaccharide modified with a basic group. In this medicine, the basic group is a group bearing a polyamine. This food contains the medicine. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a drug having a lipase inhibitory effect, a fat absorption inhibitory effect and a cholesterol absorption inhibitory effect, and a food containing this drug.

  Dietary fiber is known to have an effect of suppressing absorption of cholesterol and fat. However, although dietary fiber is highly safe, it is not effective by itself, and a large amount of intake is required to increase the absorption of cholesterol and fat. However, for example, dietary fiber such as alginic acid has a very low solubility in water and gels, making it difficult to take a large amount. In addition, excessive intake of dietary fiber may cause adverse effects such as suppression of mineral absorption in the body.

It is known from animal experiments that chitosan, which is a kind of dietary fiber, is superior in fat digestion absorption suppression action compared to other dietary fibers (see, for example, Non-Patent Document 1).
However, since chitosan hardly dissolves in water, it must be solubilized using an organic acid such as lactic acid or acetic acid when ingested as a liquid such as a beverage. Moreover, since chitosan has an intense savory taste, it is difficult to blend it into a beverage as it is. Furthermore, since chitosan and lipid polymers are formed when chitosan is taken, feces become hard and possible side effects such as frequent constipation have been pointed out.

  As a method for lowering blood cholesterol level, there has been conventionally known a therapy in which a basic anion exchange resin in which an aliphatic quaternary ammonium salt is fixed as a functional group to a styrene resin is orally administered to adsorb bile acids. (For example, Patent Documents 1, 2, and 3). However, this therapy has a drawback that it is very difficult to drink because the daily dose is as high as 8 to 16 g / day and the anion exchange resin does not dissolve in water.

US Pat. No. 3,499,960 US Pat. No. 3,780,171 Japanese Examined Patent Publication No. 61-54457 Osamu Kanai, “Chemistry and Biology”, 34, 553-557 (1996)

  The present invention is excellent in lipase inhibitory effect, fat absorption inhibitory effect, and cholesterol absorption inhibitory effect, has a little bitter taste, is easy to be added to foods and processed, has a bile acid reabsorption inhibitory effect, and is highly safe. It is an object of the present invention to provide an easily ingestible drug and a food having a lipase inhibitory effect, a fat absorption inhibitory effect, and a cholesterol absorption inhibitory effect containing the drug.

  As a result of diligent research, the present inventors have found that the above problems can be solved by a drug containing a polysaccharide modified with a basic group and a food containing the drug, and have completed the present invention based on this finding. .

The present invention is constituted by the following.
(1) A drug containing a polysaccharide modified with a basic group.
(2) The drug according to (1) above, wherein the basic group is a group containing a polyamine.
(3) The drug according to (1), wherein the basic group is a group containing a tertiary amine.
(4) The drug according to (1) above, wherein the basic group is a group containing a quaternary ammonium salt.
(5) The drug according to any one of (1) to (4), wherein the polysaccharide is an indigestible polysaccharide.
(6) The drug according to any one of (1) to (5), wherein the polysaccharide is a water-soluble polysaccharide.
(7) A food containing the drug according to any one of (1) to (6).

  The drug of the present invention is excellent in lipase inhibitory effect, fat absorption inhibitory effect and cholesterol absorption inhibitory effect, has little pungency, is easy to add to foods and is processed, has a bile acid reabsorption inhibitory effect, and is safe By selecting a polysaccharide and a basic group, the solubility in water, the viscosity when dissolved in water, the effect of suppressing fat absorption, and the like can be appropriately adjusted. Moreover, the foodstuff which added this chemical | medical agent shows the outstanding lipase inhibitory effect, fat absorption inhibitory effect, and cholesterol absorption inhibitory effect.

The drug having a lipase inhibitory effect, a fat absorption inhibitory effect and a cholesterol absorption inhibitory effect of the present invention comprises a polysaccharide modified with a basic group (hereinafter referred to as a base-modified polysaccharide).
Specific examples of the polysaccharide used in the present invention include agarose, agaropectin, gum arabic, alginic acid, carrageenan, callose, xylan, guar gum, keratan sulfate, dermatan sulfate, dextrin, schizophyllan, zebra palm mannan, konjac mannan, arabinan, cellulose , Polysaccharides extracted from natural products such as tamarind seed gum, tragagant gum, pustulan, fucoidan, funolan, porphyran, pectin, locust bean gum, starch, nigellan, lichenan, lentinan, propylene glycol alginate, ethyl cellulose, hydroxyethyl cellulose, hydroxy Examples thereof include chemically modified / synthetic polysaccharides such as propylcellulose, polydextrose, and modified starch.

  In particular, when the polysaccharide is an indigestible polysaccharide, a drug obtained by modifying the polysaccharide with a basic group can be expected to promote bowel movement as a dietary fiber, improve bowel regulation such as constipation improvement, and prevent colon cancer. Examples of indigestible polysaccharides include agarose, agaropectin, gum arabic, alginic acid, carrageenan, carose, xylan, guar gum, keratan sulfate, dermatan sulfate, schizophyllan, elephant palm mannan, konjac mannan, arabinan, cellulose, tamarind seed gum, Polysaccharides extracted from natural products such as tragagant gum, pustulan, fucoidan, funolan, porphyran, pectin, locust bean gum, nigeran, lichenan, lentinan, propylene glycol alginate, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, polydextrose, processed Examples thereof include chemically modified and synthetic polysaccharides such as starch.

  In addition, when the polysaccharide is a water-soluble polysaccharide, the drug obtained by modifying it with a basic group is often water-soluble and is suitably used for foods such as various beverages, soups, stews, and solutions. it can. Examples of water-soluble polysaccharides include agarose, agaropectin, gum arabic, alginic acid, carrageenan, carose, guar gum, keratan sulfate, dermatan sulfate, dextrin, schizophyllan, tamarind seed gum, tragagant gum, funolan, porphyran, locust bean gum, nigeran, lichenan And polysaccharides extracted from natural products such as lentinan, and chemically modified / synthetic polysaccharides such as propylene glycol alginate, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, and polydextrose.

  Specific examples of basic groups for modifying polysaccharides include aminoalkyl groups such as aminoethyl group and aminopropyl group, alkylaminoalkyl groups such as dimethylaminoethyl group, aminophenyl group, dimethylaminophenyl group, and aminobenzyl group. An aromatic ring-containing amino group, an aliphatic ring-containing amino group such as a piperidino group, a piperidyl group or a pyrrolidino group, a group containing a quaternary ammonium salt such as a trimethylammoniopropyl group or a triethylammoniopropyl group, N- (2 Examples include groups containing diamines such as-(diethylamino) ethyl) aminoethyl groups, groups containing polyamines such as diethylenetriamine, triethylenetetramine, polyallylamine, polyethyleneimine, polyvinylpyridine, α-polylysine, and ε-polylysine. . These may be used alone or in combination of two or more.

Above all, if the basic group is a group containing polyamine, the substitution rate of the basic group can be changed greatly, and the lipase inhibitory effect, fat absorption inhibitory effect, cholesterol absorption inhibitory effect, etc. of the obtained drug can be adjusted in a wide range. Is possible. Furthermore, polysaccharides modified with ε-polylysine have high safety and can be suitably used for the drug of the present invention.
Also, if the basic group is a group containing a tertiary amine, the tertiary amine has a strong basicity, so even if the substitution rate of the basic group is small, the resulting drug has a large lipase inhibitory effect and fat absorption suppression The effect and the cholesterol absorption inhibitory effect can be expected.
Furthermore, if the basic group is a group containing a quaternary ammonium salt, the basic agent is stronger than the tertiary amine possessed by the quaternary ammonium salt. Can be expected to have a large lipase inhibitory effect, fat absorption inhibitory effect, and cholesterol absorption inhibitory effect.

  The method for modifying the polysaccharide with a basic group is not particularly limited, but for example, the methods described in US Pat. No. 2813093, US Pat. No. 2,872,217, US Pat. No. 2,970,140 and the like can be used.

  The drug of the present invention may be administered as it is, but as long as it is necessary and does not impair the object of the present invention, together with pharmaceutically acceptable additives, solutions, suspensions, powders, granules, You may administer in the form of a tablet, a capsule, etc. Additives include sugars such as lactose, sucrose, and glucose, inorganic substances such as calcium carbonate and calcium sulfate, starch, crystalline cellulose, distilled water, purified water, sesame oil, soybean oil, corn oil, olive oil, cottonseed oil, etc. Mention may be made of what is being used. In formulating, additives such as binders, lubricants, dispersants, suspending agents, emulsifiers, diluents, buffers, antioxidants, and bacterial inhibitors can be used.

  It is also effective to administer the drug of the present invention in food. Examples of the food containing the agent of the present invention include bread, pasta, udon, cake, cookies, biscuits, hamburger, meat sauce, dumplings, sausage, curry, stew, jam, chocolate, corn soup and the like. In order to contain the drug of the present invention in the food, there can be exemplified a method of uniformly mixing the dough in the production process of the food dough, and a method of uniformly dissolving in the food in the case of a liquid food. . Although the content rate of the chemical | medical agent of this invention in these foodstuffs is suitably changed with the kind and intake of a foodstuff, 1-20 weight% can be illustrated.

Dietary fiber can be used in combination with the agent of the present invention. Dietary fiber that can be used in combination is not particularly limited, but pectin, polydextrose, alginic acid, sodium alginate, galactomannan, glucomannan, tamarind seed gum, xanthan gum, gellan gum, carrageenan, caraya gum, carboxymethylcellulose, cellulose, hemicellulose, lignin, insoluble pectin And apple fiber, sugar beet fiber, corn fiber, wheat fiber, soybean fiber, chitin and the like, which are composite dietary fibers. These dietary fibers may be used independently and may be used in combination of 2 or more types as appropriate.
Among them, preferable dietary fibers include apple fiber, sugar beet fiber, corn fiber, wheat fiber, and soybean fiber.

Below, the compounding example of the foodstuff containing the chemical | medical agent of this invention is shown. In addition, the number in a compounding example shows a weight part.
Food formulation example 1 (spaghetti dough)
Flour 100 parts by weight Water 30
Agent of the present invention 0.1-0.2

Food formulation example 2 (bread dough)
Flour 100 parts by weight Water 64
East 3
Salt 2
Sugar 6
Nonfat dry milk 3
Oils and fats 5
Agent of the present invention 0.1-0.2

Food formulation example 3 (hamburger dough)
40 parts by weight ground beef tallow 5
Onion 20
Raw bread crumbs 8
Starch 8
Water 5
Spices & seasonings 1.2
Agent of the present invention 0.2-0.5

EXAMPLES Hereinafter, although it demonstrates still in detail according to an Example, this invention is not limited to these Examples. The degree of substitution of basic groups was indicated by the number of basic groups introduced per saccharide unit. Moreover, the following% display means weight% unless there is particular description.
1. Synthesis of base-modified polysaccharide diethylaminoethyl group-substituted polydextrose (DEAE-PD) (Synthesis Example 1)
To a 1000 ml three-necked flask equipped with a thermometer, a dropping funnel, and a stirrer, 40 g of NaOH and 170 ml of ion exchange water were charged and dissolved uniformly. While cooling to 10 ° C. in an ice bath, 60.0 g of polydextrose (manufactured by Wako Pure Chemical Industries, Ltd.) was added and dissolved uniformly. After 30 minutes, the ice bath was removed, and 35 g of diethylaminoethyl chloride hydrochloride dissolved in 45 ml of ion exchange water was added dropwise in several portions. After completion of the dropping, the temperature was raised to 80 ° C. and reacted at 80 ° C. for 35 minutes.
After completion of the reaction, the reaction vessel was ice-cooled to room temperature and neutralized with a 6 mol / l-HCl aqueous solution. Dialyzed with ion-exchanged water and lyophilized to obtain diethylaminoethyl group-substituted polydextrose (DEAE-PD). The yield was 45.0g.

2. Synthesis of base-modified polysaccharide DEAE-PD (Synthesis Examples 2 to 5)
A 200 ml three-necked flask equipped with a thermometer, a dropping funnel, and a stirrer was charged with the amount of polydextrose (manufactured by Wako Pure Chemical Industries, Ltd.) and ion-exchanged water described in Table 2 and dissolved uniformly. . While cooling to 10 ° C. with an ice bath, NaOH dissolved in the amount of ion-exchanged water shown in Table 2 was added dropwise over 10 minutes. Twenty minutes after the completion of dropping, the ice bath was removed, and diethylaminoethyl chloride hydrochloride dissolved in the amount of ion-exchanged water shown in Table 2 was added dropwise over 5 minutes. 30 minutes after the completion of dropping, the temperature was raised to 80 ° C., and the reaction was carried out at 80 ° C. for 3 hours.
After completion of the reaction, the reaction vessel was ice-cooled to room temperature, and the reaction mixture was poured into 300 ml of methanol to precipitate a polymer. The polymer was filtered, dissolved again in 20 ml of ion-exchanged water, poured into 250 ml of methanol, and the polymer was precipitated.
The polymer filtered off was dialyzed with ion-exchanged water and freeze-dried to obtain diethylaminoethyl group-substituted polydextrose (DEAE-PD). Yields were Synthesis Example 2: 0.87 g, Synthesis Example 3: 0.39 g, Synthesis Example 4: 0.15 g, and Synthesis Example 5: 1.07 g.

3. Substitution degree measurement of base-modified polysaccharide DEAE-PD 0.004 g of the obtained DEAE-PD was dissolved in 0.8 g of heavy water and measured by ¹ H-WEFT-NMR method. The degree of substitution of the basic group was calculated from the area ratio of low magnetic field proton (near 2.4-5.5 ppm) / high magnetic field proton (near 0.5-1.5 ppm). The degree of substitution of DEAE-PD in Synthesis Examples 1 to 5 is shown in Table 1.

(Table 1)

4). Lipidase inhibitory effect test 4-1. Preparation of substrate solution Triolein 80 mg, phosphatidylcholine 10 mg, and sodium taurocholate 5 mg were mixed with 9 ml of 0.1 mol / l-NaCl-0.1 mol / l-TES buffer (pH 7.0) and sonicated. Emulsification gave a substrate solution.
4-2. Preparation of Base Modified Polysaccharide DEAE-PD Solution 1 ml of 0.1 mol / l-NaCl-0.1 mol / l-TES buffer (pH 7.0) was added to the base modified polysaccharide obtained in Synthesis Examples 1, 2, 3 and 5. Dissolved to prepare various concentrations of DEAE-PD solutions.

4-3. Preparation of Lipolytic Enzyme Solution As the pancreatic lipase, an enzyme purified from the rat pancreas by electrophoresis was used. It was diluted to 4 μg / ml with 0.1 mol / l-NaCl-0.1 mol / l-TES buffer (pH 7.0) and used.

4-4. Measurement of lipolytic enzyme inhibitory activity To 100 μl of the substrate solution prepared above, 100 μl of the base-modified polysaccharide DEAE-PD solution prepared above was added and shaken for 5 minutes, and then 50 μl of the lipolytic enzyme solution prepared above was added at 37 ° C. Shake for 30 minutes. Next, 3 ml of an extraction solvent (chloroform: methanol: heptane = 49: 2: 49) was added and shaken for 10 minutes, followed by centrifugation at 2500 rpm for 5 minutes. After removing the upper layer liquid with an aspirator, a copper reagent (0.1 mol / l-triethanolamine-0.06 mol / l-NaOH-0.05 mol / l-copper nitrate trihydrate solution 200 ml was added to the lower layer liquid. 1 ml of sodium chloride dissolved in 66 g) was shaken for 10 minutes, and then centrifuged at 2500 rpm for 10 minutes. Next, 0.5 ml of the upper layer solution was collected, and 0.5 ml of a coloring reagent (0.1 w / v% -vasocproin, 0.05 w / v% -butylated hydroxyanisole-chloroform solution) was added and shaken, and the wavelength was 480 nm. The amount of free fatty acid produced, that is, the activity of the lipolytic enzyme was examined. The activity of lipolytic enzyme is shown in FIG.

[Measurement of lipolytic enzyme activity]
Polydextrose (manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved in 1 ml of 0.1 mol / l-NaCl-0.1 mol / l-TES buffer (pH 7.0) to prepare polydextrose solutions of various concentrations. Instead of the base-modified polysaccharide solution, 100 μl of polydextrose solution of various concentrations was added to 100 μl of the substrate solution prepared in the same manner as in Example 1, and the lipolytic enzyme inhibitory activity was measured in the same manner as in Example 1. The results are shown in FIG.

[Synthesis of Base-Modified Polysaccharide Piperidinoethyl Group-Substituted Polydextrose (Synthesis Examples 6-9)]
A 200 ml three-necked flask equipped with a thermometer, a dropping funnel, and a stirrer was charged with the amount of polydextrose (manufactured by Wako Pure Chemical Industries, Ltd.) and ion-exchanged water described in Table 2 and dissolved uniformly. . While cooling to 10 ° C. with an ice bath, NaOH dissolved in the amount of ion-exchanged water shown in Table 2 was added dropwise over 10 minutes. 20 minutes after the completion of dropping, the ice bath was removed, and 1- (2-chloroethyl) piperidine hydrochloride dissolved in the amount of ion-exchanged water shown in Table 2 was added dropwise over 5 minutes. 30 minutes after the completion of dropping, the temperature was raised to 80 ° C., and the reaction was carried out at 80 ° C. for 3 hours.
After completion of the reaction, the reaction vessel was ice-cooled to room temperature, neutralized with hydrochloric acid, dialyzed with ion exchange water, and lyophilized to obtain piperidinoethyl group-substituted polydextrose (PIPE-PD). Yields were: Synthetic Example 6: 0.71 g, Synthetic Example 7: 0.50 g, Synthetic Example 8: 0.53 g, and Synthetic Example 9: 1.11 g.
The obtained base-modified polysaccharide PIPE-PD was subjected to substitution degree measurement and lipolytic enzyme inhibitory effect test in the same manner as in Example 1. The results are shown in Table 2 and FIG.

(Table 2)

1. Synthesis of base-modified polysaccharide 3-triethylammonio-2-hydroxypropyl group-substituted corn starch (Synthesis Examples 10 to 12)]
A 300 ml three-necked flask equipped with a thermometer and a stirrer was charged with 20.2 g of triethylamine, 18.5 g of epichlorohydrin and 100 ml of ion-exchanged water, and stirred at room temperature for 5 hours. Next, the reaction mixture was distilled off under reduced pressure at 30 ° C. using an evaporator to obtain triethylglycidyl ammonium chloride. Yield 32.8g. Triethyl glycidyl ammonium chloride was used in the following reaction without purification.
A 200 ml three-necked flask equipped with a thermometer, a dropping funnel, and a stirrer was charged with the amount of corn starch (manufactured by Kawamitsu Bussan Co., Ltd.), sodium chloride and ion-exchanged water as shown in Table 4 while stirring. The amount of triethylglycidylammonium chloride obtained above in the amount of 4 was charged, followed by the amount of 10% sodium hydroxide aqueous solution in the amount of Table 4. After stirring at room temperature for 2 hours and 30 minutes, the temperature was raised to 35 ° C. with an oil bath and the reaction was carried out at 35 ° C. for 68 hours.
After completion of the reaction, the reaction mixture was acidified with 2.4M hydrochloric acid and then filtered, then twice with 0.07 mol / l-hydrochloric acid, twice with 0.2% aqueous sodium hydroxide, twice with ion-exchanged water, and with methanol. This was washed twice and dried at 40 ° C. for 5 hours and at 80 ° C. for 6 hours to obtain 3-triethylammonio-2-hydroxypropyl group-substituted corn starch (TEAP-ST). Yields were Synthesis Example 14: 14.4 g, Synthesis Example 15: 7.70 g, and Synthesis Example 16: 10.6 g.

2. Substitution degree measurement of base-modified polysaccharide TEAP-ST 1.000 g of base-modified polysaccharide TEAP-ST was weighed, 50 ml of 0.1 mol / l-HCl aqueous solution was added, and the mixture was stirred for 2 hours and allowed to stand until the next day. 10 ml of the supernatant was weighed and titrated with 0.1 mol / l-NaOH aqueous solution using phenolphthalein as an indicator. The degree of substitution of the basic group was calculated from the volume of 0.1 mol / l-NaOH aqueous solution required for neutralization. The results are shown in Table 3.
Further, the obtained base-modified polysaccharide was subjected to a lipolytic enzyme inhibitory effect test in the same manner as in Example 1. The results are shown in FIG.

(Table 3)

[Synthesis of Base-Modified Polysaccharide ε-Polylysine Substituted Cellulose (Synthesis Examples 13 to 15)]
In a 500 ml separable flask equipped with a thermometer, a dropping funnel, and a stirrer, porous spherical cellulose (water content 88.8%) obtained by the method described in JP-A-10-195103 in the amount described in Table 5 was used. ) And ion-exchanged water were added, and a 20% aqueous solution of sodium hydroxide described in Table 5 was added while stirring. The temperature was raised to 30 ° C. in an oil bath, and after stirring for 1 hour, the amount of epichlorohydrin shown in Table 5 was added and reacted at 30 ° C. for 2 hours.
After completion of the reaction, the cellulose was collected by filtration, washed 5 times with ion exchange water, confirmed to be neutral, and returned to the separable flask again. The amounts of ion-exchanged water and 25% -ε-polylysine aqueous solution (manufactured by Chisso Corp.) described in Table 5 were added thereto, heated to 45 ° C. with an oil bath, and reacted for 2 hours.
After completion of the reaction, the cellulose was collected by filtration, washed 5 times with ion-exchanged water and twice with methanol, dried at 60 ° C. for 1 hour, 80 ° C. for 2 hours, and 100 ° C. for 3 hours, and ε-polylysine substituted cellulose ( EPL-CEL) was obtained. Yields were Synthesis Example 17: 7.90 g, Synthesis Example 18: 8.55 g, Synthesis Example 19: 7.21 g.
About the obtained base modification polysaccharide, the substitution degree was measured by the method similar to Example 4, and the lipolytic enzyme inhibitory effect test was implemented by the method similar to Example 1. FIG. The results are shown in Table 4 and FIG.

(Table 4)

1. Synthesis of base-modified polysaccharide aminobenzoyl group-substituted polydextrose (Synthesis Example 16)
A 100 ml three-necked flask equipped with a thermometer, a nitrogen gas inlet tube and a stirrer is charged with 6.0 g of polydextrose (manufactured by Wako Pure Chemical Industries, Ltd.) and 20 ml of dimethylformamide and heated to 40 ° C. in an oil bath. And uniformly dissolved. To this, 5.0 g of triethylamine was added, and 13.52 g of p-nitrobenzoyl chloride dissolved in 16 ml of dimethylformamide was added dropwise over 30 minutes while maintaining 40 ° C. 50 minutes, 3 hours, and 5 hours after the completion of dropping, 1.5 g of triethylamine was added, and the reaction was performed for 6 hours.
After completion of the reaction, the reactor was allowed to cool to room temperature, and the reaction solution was poured into 300 ml of isopropanol to precipitate the polymer and filtered. The polymer was suspended again in 200 ml of isopropanol heated to 40 ° C., stirred for 1 hour, filtered, and dried at 80 ° C. for 4 hours to obtain nitrobenzoyl group-substituted polydextrose.
In a 100 ml three-necked flask equipped with a hydrogen gas introduction tube and a stirrer, the entire amount of the nitrobenzoyl group-substituted polydextrose obtained above, 30 ml of tetrahydrofuran, 5 ml of ion-exchanged water and 10% -palladium-carbon (water content 51.4%) 0.4 g was charged and the inside of the reaction vessel was replaced with hydrogen gas, followed by stirring at room temperature for 8 hours.
After completion of the reaction, the catalyst was removed by filtration, concentrated to about 20 ml, poured into 200 ml of isopropanol, the polymer was precipitated and filtered. It dried at 80 degreeC for 6 hours, and aminobenzoyl group substituted polydextrose (ABZ-PD) was obtained. The yield was 4.8g.

2. Substitution degree measurement of base-modified polysaccharide ABZ-PD 0.004 g of the obtained ABZ-PD was dissolved in DMSO-d6 / heavy water = 12/1 (wt / wt) 0.8 g, and measured by ¹ H-WEFT-NMR method. did. The degree of substitution of the basic group was calculated from the area ratio of aromatic protons (around 7.5-9.5 ppm) / non-aromatic protons (around 2.4-5.5 ppm). The degree of substitution calculated in this way was 0.59.
Furthermore, the result of the lipolytic enzyme inhibitory effect test conducted by the same method as in Example 1 is shown in FIG.

[Synthesis of base-modified polysaccharide diethylaminoethylamide group-containing pectin (Synthesis Example 17)]
A 100 ml three-necked flask equipped with a thermometer and a stirrer was charged with 1.76 g of pectin (manufactured by Tokyo Chemical Industry Co., Ltd.), 50 ml of ion-exchanged water and 1.16 g of N, N-diethylethylenediamine, and dissolved uniformly. It was. To this was added 22 ml of 1 mol / l-HCl, followed by 1.92 g of 1-ethyl-3- (N, N-dimethylaminopropyl) carbodiimide, and the mixture was stirred at room temperature for 3 days. After adding 1 ml of 1 mol / l-HCl, dialyzed with ion exchange water, 0.1% -NaOH aqueous solution and ion exchange water in that order and freeze-dried to obtain diethylaminoethylamide group-containing pectin (DEEDA-PEC). The yield was 0.68g. The degree of substitution measured by the same method as in Example 1 was 0.54. Furthermore, the result of the lipolytic enzyme inhibitory effect test carried out by the same method as in Example 1 is shown in FIG.

[Synthesis of Base-Modified Polysaccharide Diethylaminoethylamide Group-Containing Pectin (Synthesis Example 18)]
A 100 ml three-necked flask equipped with a thermometer and a stirrer was charged with 1.76 g of pectin (manufactured by Tokyo Chemical Industry Co., Ltd.), 50 ml of ion-exchanged water and 0.349 g of N, N-diethylethylenediamine and dissolved uniformly. It was. To this, 5 ml of 1 mol / l-HCl, 0.575 g of 1-ethyl-3- (N, N-dimethylaminopropyl) carbodiimide and 15 ml of ion-exchanged water were added, and the mixture was stirred at room temperature for 3 days. Dialysis was performed in the order of ion-exchanged water, 0.1% -NaOH aqueous solution, and ion-exchanged water, and lyophilized to obtain diethylaminoethylamide group-containing pectin (DEEDA-PEC). The yield was 0.52g. Further, the degree of substitution measured by the same method as in Example 1 was 0.14. Furthermore, the result of the lipolytic enzyme inhibitory effect test carried out by the same method as in Example 1 is shown in FIG.

  From FIG. 1-6, it can be seen that a variety of base-modified polysaccharides can be obtained, from those showing lipolytic enzyme inhibitory activity at a concentration of about 1 μg / ml to those showing lipolytic enzyme inhibitory activity on the order of several mg / ml. That is, it was shown that the lipolytic enzyme inhibitory activity can be controlled by changing the type of polysaccharide, the type of basic substituent and the substitution rate.

[Taste test]
A taste test was conducted by five people using the base-modified polysaccharide synthesized in Synthesis Examples 2, 8, and 17 and chitosan (chitosan 10 manufactured by Wako Pure Chemical Industries, Ltd.) as a comparative example. The base-modified polysaccharide synthesized in Synthesis Examples 2, 8, 10, and 21 was used as a 1% aqueous solution, and chitosan was mixed with lactic acid at 2/1 (wt / wt) and used as a 1% aqueous solution in the test. The evaluation of bitterness and gummy taste was expressed as “feel bitter and gummy”, “feel slightly bitter and gummy” and “do not feel bitter and gummy”. The results are shown in Table 5. The numbers in the table indicate the number of people.
As a result of the test, it was shown that the base-modified polysaccharide of the present invention has less bitterness and bitterness than the chitosan as a comparative example and is easy to take. Moreover, this result shows that the addition to a foodstuff is easy.

(Table 5)

[Solubility test]
Base modified polysaccharides synthesized in Synthesis Examples 5 and 9, as Comparative Example 2, Wako Pure Chemical Industries, Ltd. Chitosan 10, Chitosan 100, and sodium alginate (Kimitsu Chemical Industries, Ltd. Kimitsu Algin I-3) in pure water Dissolved to make a 1% aqueous solution. Since chitosan 10 and chitosan 100 do not dissolve in pure water, 1% solution was prepared by dissolving in 0.5% -acetic acid aqueous solution. Table 6 shows the viscosity of each aqueous polysaccharide solution.
As a result of the test, the base-modified polysaccharide of the present invention is easily dissolved in pure water without adjusting the pH as compared with chitosan as a comparative example, and dissolved in water as compared with alginic acid as a comparative example. It was shown that the viscosity was low and it was easy to take a large amount. Moreover, this result shows that the addition to a foodstuff is easy.

(Table 6)

[Triglyceride absorption inhibition test]
Corn oil (6 ml), cholic acid (80 mg), cholesterol oleate (2 mg), and pure water (6 ml) were mixed, sonicated and emulsified to obtain a corn oil emulsion. Male rats weighing 200 g were fasted overnight, divided into two groups, and the control group was administered with 1 ml of pure water added to 1 ml of corn oil emulsion. In the DEAE-administered group (DEAE-PD group), 1 ml of a DEAE-PD aqueous solution (DEAE-PD obtained in Synthesis Example 1 dissolved in pure water to a concentration of 50 mg / ml) is added to 1 ml of corn oil emulsion. In addition, it was administered. Blood was collected over time from the tail vein or the tail artery, and the triglyceride content was measured. FIG. 7 shows the blood triglyceride concentration after administration. The blood triglyceride concentration increased significantly 2 hours after administration when DEAE-PD was not administered (control group). However, when DEAE-PD was administered (DEAE-PD group), the blood triglyceride concentration was almost constant even after administration. It was confirmed that it did not rise.

[Inhibition of body weight gain, serum lipid elevation and adipose tissue weight increase in mice fed with high fat diet]
C57BL / 6 male mice (7 weeks old) were purchased, and after 1 week of preliminary breeding, they were raised for 20 weeks on the following high fat diet. One group used 8 animals. In the DEAE-PD administration group (DEAE-PD group), DEAE-PD obtained in Synthesis Example 1 was administered twice in the morning and evening so as to be 500 mg / kg, and the control group administered the same amount of pure water. Body weight was measured over time. In addition, serum lipids (triglycerides and cholesterol) and adipose tissue weight at 20 weeks were measured.
FIG. 8 shows changes in the body weight of mice administered with a high fat diet. The DEAE-PD group showed a tendency to lose weight from 10 weeks compared to the control group.
FIG. 9 shows the serum lipid content at 20 weeks. A decrease in serum triglyceride and cholesterol content was observed with DEAE-PD administration.
FIG. 10 shows the adipose tissue weight at 20 weeks. A tendency to decrease the weight of adipose tissue was observed with DEAE-PD administration.
・ Composition of high fat food (per 100g)
Unsalted butter (milk fat) 45.0g
Corn starch 17.1g
Sucrose 10.0g
Casein 20.0g
Cellulose powder 3.0g
Mineral mix 3.5g
Vitamin mix 1.0g
Choline chloride 0.4g
Total 100g

  It can be used for therapeutic drugs for hyperlipidemia based on lipase inhibitory effect, fat absorption inhibitory effect and cholesterol absorption inhibitory effect, food additives for preventing hyperlipidemia and obesity, and health foods.

Test of inhibitory effect of lipolytic enzyme by DEAE-PD (Example 1) Lipidase inhibitory effect test with PIPE-PD (Example 2) Lipidase inhibitory effect test by TEAP-ST (Example 3) Lipidase inhibitory effect test by EPL-CEL (Example 4) Lipidase inhibitory effect test with ABZ-PD (Example 5) Lipidase inhibitory effect test with DEEDA-PEC (Example 6) Effect of DEAE-PD on triglyceride absorption (Example 9) Body weight of mice administered with high fat diet (Example 10) Effect of serum lipids (Example 10) Adipose tissue weight of high fat diet administered mice (Example 10)

Claims (7)

  A drug consisting of a polysaccharide modified with a basic group.   The drug according to claim 1, wherein the basic group is a group containing a polyamine.   The drug according to claim 1, wherein the basic group is a group containing a tertiary amine.   The drug according to claim 1, wherein the basic group is a group containing a quaternary ammonium salt.   The drug according to any one of claims 1 to 4, wherein the polysaccharide is an indigestible polysaccharide.   The drug according to any one of claims 1 to 5, wherein the polysaccharide is a water-soluble polysaccharide. The foodstuff containing the chemical | medical agent of any one of Claims 1-6.

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