JP2003012536A - Lipase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new lipase inhibitor containing as an active ingredient a new excellent lipase-inhibiting action-having substance found from natural matters having confirmed safety. SOLUTION: This lipase inhibitor is characterized by containing an Ampelpsis cantoniensis extract and/or an Engelhardtia chrysolepis H. extract containing dihydroflavonol compounds as an active ingredient.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel lipase inhibitor. More specifically, it suppresses the generation of malodor caused by fats and oils present in the living environment such as food and drink, cosmetics, daily sundries, etc. through lipase inhibitory action, and makes it possible to directly contact the skin such as cosmetics and daily sundries. The present invention relates to a lipase inhibitor useful for suppressing rough skin caused by existing fats and oils, and for suppressing obesity caused by fats and oils present in food and drink.

[0002]

BACKGROUND ART Oils and fats contained in foods and drinks are hydrolyzed by lipases produced by microorganisms existing in the environment to produce glycerol and fatty acids. The produced fatty acid is oxidized to generate aldehydes and the like, which causes bad odor and deterioration, which is a big problem in maintaining the quality of foods and drinks.

In the skin, fatty acids are produced from sebum by the action of lipase produced by microorganisms present in the skin or lipase of epidermal keratinocytes, and the produced fatty acids activate and activate neutrophils. Neutrophils release active oxygen, which causes acne and dermatitis.

On the other hand, lipase, which is a kind of digestive enzyme,
Digestion and absorption are promoted by decomposing fats and oils in the food and drink that have been eaten and drank. Therefore, it is considered that obesity, hyperlipidemia, atherosclerosis and arteriosclerosis can be treated or prevented by inhibiting the action of lipase.

Examples of the lipase inhibitor which inhibits the activity of lipase which adversely affects foods and beverages, cosmetics, skin, etc., include, for example, lemongrass, allspice, cinnamon, cloves, Assenyaku, guava, and medussweet. , A lipase inhibitor using an extract from a plant such as Cola de Cavallo, Loquat, Dokatsu, Ryokyo, Areca, Euphorbia, Sunpens, Ketsumeishi, etc. has been proposed (JP 2001-120237 A, JP 2).
000-103741, JP-A-11-228338, JP-A-10-265364, JP-A-5-255100),
At present, what has been satisfactory in terms of effectiveness and safety has not yet been developed.

[0006]

Therefore, the present invention has found a substance having an excellent lipase inhibitory action among natural substances of which safety has been confirmed, and a novel lipase inhibitor containing the substance as an active ingredient. The purpose is to provide.

[0007]

In order to achieve the above object, the first lipase inhibitor provided by the present invention is
It is characterized by containing a wisteria tea extract and / or a yellow-fruit extract as an active ingredient. In the first lipase inhibitor of the present invention, the wisteria tea extract is an extract obtained by extracting the wisteria tea branches and leaves with a polar solvent, and the yellow mud extract is the yellow mud branches and leaves. It is preferably an extract obtained by extracting with a polar solvent.

In order to achieve the above object, the second lipase inhibitor provided by the present invention is characterized by containing dihydroflavonols as an active ingredient. In the second lipase inhibitor of the present invention, the dihydroflavonols are preferably ampelopsin, astilbin, neoastilbin, neoisoastilbin or isoastilbin.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, the “Fujicha extract” and the “yellow mulberry extract” refer to the extracts obtained by using Fuji tea and yellow mulberry as raw materials for extraction, respectively. Is an extract obtained by using Fuji tea or yellow mullet as an extraction raw material, a dilute solution or a concentrated solution of the extract, a dried product obtained by drying the extract, or a crudely purified product or a purified product thereof. Is included.

Fuji tea (scientific name: Ampe
lopsis grossedentata (Hand.-Mazz.) WTWang, or A
mpelopsis cantoniensis (Hook.et Arn.) Panch.) is a perennial climbing plant belonging to the vine family. In addition to being used as a beverage, the root part or whole plant of Fuji tea is used as a folk medicine for the treatment of jaundice hepatitis, colds, sore throat, acute conjunctival inflammation, etc., It is a highly safe plant. Wisteria tea grows naturally in large areas from central to southern China and Taiwan and is readily available from these areas. As the extraction raw material, whole plants, branches, leaves, branches, etc. of Fuji tea can be used, but the branches and leaves are preferably used.

Yellow husk (scientific name: Enge used as an extraction raw material
lhardtia chrysolepis Hance) is an evergreen tree plant belonging to the family Walnut. Yellow mud is a highly safe plant that has been used as a kind of sweet tea since ancient times. Huangshu grows naturally in regions from southern China to Taiwan and is readily available from these regions. As the extraction raw material, the branches, leaves, branches and the like of yellow mullet can be used, but the branches and leaves are preferably used.

[0012] Fuji tea and / or yellow musk as an extraction raw material can be used as it is or after being dried and then pulverized as it is or by a pulverizer. Drying may be performed in the sun or using a commonly used dryer. The wisteria tea and / or yellow mud as an extraction raw material may be used after being subjected to a pretreatment such as degreasing with a nonpolar solvent such as hexane or benzene. By performing pretreatment such as degreasing, it is possible to efficiently perform the extraction treatment of the wisteria tea and / or the yellow husk with the polar solvent.

In the extraction treatment, it is preferable to use a polar solvent as the extraction solvent. The component showing a lipase inhibitory action contained in Fuji tea or yellow husk can be easily extracted by an extraction treatment using a polar solvent as an extraction solvent.

Specific examples of suitable polar solvents include water and hydrophilic organic solvents. These may be used alone or in combination of two or more kinds at a temperature from room temperature to the boiling point of the solvent. You can

Water that can be used as the extraction solvent in the present invention includes pure water, tap water, well water, mineral water, mineral water, hot spring water, spring water, fresh water, and the like, as well as those that have been subjected to various treatments. Be done. Examples of the treatment applied to water include heating, sterilization, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering and the like. Therefore, the water that can be used as the extraction solvent in the present invention also includes hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline and the like.

Examples of the hydrophilic organic solvent that can be used as the extraction solvent in the present invention include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propylene alcohol and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1 Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as 3,3-butylene glycol, propylene glycol, isopropylene glycol, and glycerin.

When a mixed solution of two or more polar solvents is used as the extraction solvent, the mixing ratio can be adjusted appropriately. For example, when a mixed liquid of water and a hydrophilic organic solvent is used, if the hydrophilic organic solvent is a lower alcohol, 1 to 90 parts by weight relative to 10 parts by weight of water, the hydrophilic organic solvent is a lower aliphatic 1 to 10 parts by weight of water for ketones
If 40 parts by weight and the hydrophilic organic solvent is a polyhydric alcohol, 10 to 90 parts by weight is preferably mixed with 10 parts by weight of water.

The extraction treatment is not particularly limited as long as the soluble component contained in Fuji tea and / or yellow musk can be eluted in the extraction solvent, and can be carried out by a conventional method. At the time of the extraction treatment, it is not necessary to employ a special extraction method, and any device can be used at room temperature or under reflux heating.

For example, the extraction raw material is put into a treatment tank filled with an extraction solvent, and the soluble components are eluted while stirring occasionally. At this time, the extraction conditions can be appropriately adjusted depending on the extraction raw material and the like. For example, when water is used as the extraction solvent, the extraction solvent amount is usually about 1 to 1000 times the extraction raw material (weight ratio), the extraction time is usually about 30 minutes to 2 hours, and the extraction temperature is usually 40 to 90 ° C. The degree. When a mixed solution of water and ethanol is used as the extraction solvent, the amount of the extraction solvent is usually about 1 to 100 times the extraction raw material (weight ratio), the extraction time is usually about 30 minutes to 2 hours, and the extraction temperature is Is usually about 40 to 80 ° C.

After the soluble components are eluted by the extraction treatment, the extraction residue is removed by filtration to obtain an extract. The obtained extract is a diluted or concentrated solution of the extract, a dried product of the extract, or a crude product or a purified product thereof, diluted according to a conventional method,
You may perform processes, such as concentration, drying, and refinement | purification.

When the obtained extract is purified for the purpose of decolorization, deodorization, activity improvement, etc., the purification method is not particularly limited, and examples of the purification method include activated carbon treatment, resin adsorption treatment, ion exchange resin, A method such as liquid-liquid countercurrent distribution may be used. Specifically, a resin adsorption purification method in which a porous resin such as SepaBeads SP-207 and Diaion HP-20 (both manufactured by Mitsubishi Chemical Co., Ltd.) and a concentrated solution are brought into contact with each other can be employed. In this case, the active ingredient adsorbed on the resin is
It can be eluted with water, ethanol or the like.

Since the wisteria tea extract and the yellow mulberry extract have unique odors and tastes, it is possible to carry out purification for the purpose of decolorization, deodorization, etc. within a range that does not reduce the physiological activity. However, since it is not used in a large amount when it is added to cosmetics or foods, there is no practical problem even if it remains unrefined.

The extract thus obtained can be used as it is as a lipase inhibitor, but it can also be used by formulating it according to a conventional method. In the case of formulation, a pharmaceutically acceptable carrier such as dextrin or cyclodextrin and any other auxiliaries can be added to facilitate storage and handling. It can be made into any dosage form such as powder, granules and tablets by formulation. At the time of formulation, other substances having a lipase inhibitory action may be added if necessary.

[0024] The lipase inhibitor may contain either one of the wisteria tea extract and the yellow mulberry extract as an active ingredient, or may contain both of them as an active ingredient. When both are contained as active ingredients, the content ratio is not particularly limited, but is usually 0.1: 99.9 to 9
9.9: 0.1 (weight ratio), preferably 5: 95-9
It is 5: 5 (weight ratio).

The lipase inhibitor of the present invention containing a Fuji tea extract and / or a yellow-fruit extract as an active ingredient can efficiently inhibit the lipase activity. Therefore, according to the lipase inhibitor of the present invention, through the lipase inhibitory action, it is possible to prevent the decomposition of oils and fats contained in foods and drinks and cosmetics, and to suppress the malodor and spoilage of foods and drinks and cosmetics. It is possible to prevent and treat acne and dermatitis through the lipase inhibitory action, and prevent and treat obesity, hyperlipidemia, atherosclerosis and arteriosclerosis through the lipase inhibiting action.

Since the lipase inhibitor of the present invention has little adverse effect on the flavor and feeling of use of the object to be added, it can be used for a wide range of objects to be added. The object to which the lipase inhibitor of the present invention is added is not particularly limited, and specific examples thereof include foods and drinks, cosmetics, oral preparations and the like.
Specific examples of food and drink to be added include soft drinks, carbonated drinks, nutritional drinks, fruit drinks, drinks such as lactic acid drinks (including concentrated liquids and preparation powders of these drinks); ice cream, ice sorbet, shaved ice. Frozen desserts such as; noodles such as buckwheat, udon, harusame, gyoza skin, Shumai skin, Chinese noodles, instant noodles; candy, candy, gum, chocolate, snacks, biscuits, jellies, jams,
Confectionery such as cream and baked goods; processed fish and livestock products such as kamaboko, ham and sausage; dairy products such as processed milk and fermented milk; salad oil, tempura oil, margarine, mayonnaise,
Oils and fats and processed foods such as shortenings, whipped creams, dressings; seasonings such as sauces and sauces; various forms of health and nutritional supplements such as tablets and granules;
Other examples include soups, stews, salads, prepared foods, pickles, and the like, and cosmetics to be added include skin cosmetics such as ointments, creams, emulsions, lotions, and packs.

The compounding amount of the Fuji tea extract and / or the yellow-branch extract in the food or drink or the cosmetic can be appropriately adjusted depending on the kind of the food or drink or the cosmetic, the physiological activity of the extract, and the like. The rate is about 0.01 to 10% by weight in the case of foods and drinks, and about 0.001 to 5% by weight in the case of cosmetics, calculated as the dry product of the unrefined standard extract. %.

Since a large amount of wisteria tea and / or yellow mud, which is a raw material for extracting the active ingredient, is present in nature and can be stably supplied, the lipase inhibitor of the present invention can be manufactured at low cost.

[0029] The wisteria tea extract and / or the yellow mulberry extract contains dihydroflavonols, and the lipase inhibitory action of the wisteria tea and / or the yellow mulberry extract is mainly exerted by the dihydroflavonols. it is conceivable that. However, the lipase inhibitory action of the wisteria tea extract and / or the yellow-fruited extract is not limited to the lipase inhibitory action by the dihydroflavonols. Fuji tea extract contains various flavonoids and other components containing amperopsin as a main component, and yellow mud extract contains taxifolin glycosides (astilbin, neoastilbin (neoa).
stilbin), isoastilbin (isoastilbin) and neoisoastilbin (neoisoastilbin) based on various flavonoids and other components are included,
It is considered that these various components are combined to exert the excellent lipase inhibitory effect of the wisteria tea extract and / or the yellow-fruited extract.

In the present invention, "dihydroflavonols" means dihydroflavonol and its derivatives, and includes the compounds represented by the general formula (I).

[Chemical 1]

In the general formula (I), R ¹ and R ³ each independently represent a hydrogen atom or an alkyl group, and R ² , R ⁴ and R ⁵ each independently represent a hydroxyl group, an alkoxy group or a sugar residue. R ⁶ , R ⁷ and R ⁸ each independently represent a hydrogen atom, a hydroxyl group, an alkoxy group or a sugar residue.

In the general formula (I), the alkyl group represented by R ¹ or R ³ is not particularly limited,
It is preferably a linear or branched alkyl group having 1 to 5 carbon atoms. Specific examples thereof include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group,
Examples thereof include an isobutyl group, an s-butyl group, a t-butyl group, an n-pentyl group, an isopentyl group, a t-pentyl group and a neopentyl group, and among them, a methyl group is particularly preferable.

In the general formula (I), R ² , R ⁴ ,
The alkoxy group represented by R ⁵ , R ⁶ , R ⁷ or R ⁸ is not particularly limited, but preferably has 1 to 1 carbon atoms.
4 is a linear or branched alkoxy group. Specific examples thereof include a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, an s-butoxy group, and a t-butoxy group. preferable.

In the general formula (I), R ² , R ⁴ ,
The sugar residue represented by R ⁵ , R ⁶ , R ⁷ or R ⁸ is not particularly limited and may be a monosaccharide or an oligosaccharide such as a disaccharide, a trisaccharide or a tetrasaccharide. May be. The oligosaccharide may be a reducing oligosaccharide or a non-reducing oligosaccharide.

Specific examples of monosaccharides include hexoses such as glucose, fructose, mannose, galactose, talose, sorbose, tagatose and psicose,
Pentose such as xylose, arabinose, ribose, lyxose, ribulose, xylulose and the like can be mentioned. The hydroxyl group of these monosaccharides may be substituted with a hydrogen atom, an amino group, etc., any of the oxygen atoms contained in the sugar may be substituted with a sulfur atom, and in the case of aldose, The hydroxymethyl group at the end of the carbon chain may be oxidized to a carboxyl group.

A sugar having a hydroxyl group replaced with hydrogen, that is,
Specific examples of the deoxy sugar include rhamnose (6-deoxymannose) in which the hydroxyl group on the 6-position carbon atom of mannose is replaced by hydrogen, and fucose (6--deoxymannose) in which the hydroxyl group on the 6-position carbon atom of galactose is replaced by hydrogen. Deoxygalactose).

Specific examples of sugars in which a hydroxyl group is substituted with an amino group, that is, amino sugars include N-acetylglucosamine, N-acetylgalactosamine, N-acetylneuraminic acid and the like.

Specific examples of sugars in which any one of oxygen atoms is replaced by a sulfur atom, that is, thiosugar, include 1-
Thio-D-glucose etc. are mentioned.

Specific examples of the sugar in which the hydroxymethyl group at the terminal of the carbon chain of aldose is oxidized to a carboxyl group, that is, uronic acid, include glucuronic acid, galacturonic acid, mannuronic acid, iduronic acid and the like.

Specific examples of oligosaccharides include those in which one or more types of monosaccharide molecules are glycosidic linked. For example, specific examples of disaccharides include xylobiose, trehalose, levanbiose, chitobiose, 2
Homodisaccharides such as -β-glucuronosyl glucuronic acid, sucrose, lactose, vicyanose, sambubiose, melibiose, epicellobiose, turanose, lactulose and other heterosugars, and specific examples of trisaccharides include raffinose and umbellife. Examples thereof include sucrose and the like. Specific examples of the tetrasaccharide include sucrose and the like.

The monosaccharide molecule constituting the oligosaccharide may be deoxy sugar, methyl sugar, amino sugar, thio sugar, uronic acid, etc. For example, specific examples of the dioxy sugar-containing disaccharide include rutinose and robinobiose. And so on,
Specific examples of disaccharides containing amino sugars include trehalosamine and the like, and specific examples of disaccharides containing uronic acid include cellobiouronic acid and the like.

In the general formula (I), R ² , R ⁴ ,
The binding mode of the sugar residue represented by R ⁵ , R ⁶ , R ⁷ or R ⁸ and the non-sugar moiety is not particularly limited, but normally, the sugar residue is bonded to the non-sugar moiety through the hydroxyl group of the non-sugar moiety. It has an ether bond.
Other bonding modes include, for example, thioglucoside bond.

The dihydroflavonols contained in the Fuji tea extract and exhibiting lipase inhibitory activity include amperopsin (in the general formula (I), R ¹ and R ³ are hydrogen atoms, and R ² , R ⁴ and R ^{5 are} , R ⁶ , R ⁷ and R ⁸ are hydroxyl groups). Further, as dihydroflavonols contained in the yellow mulberry extract and exhibiting lipase inhibitory activity, astilbin which is a taxifolin glycoside (in the general formula (I), R ¹ , R ³ and R ⁸ are hydrogen atoms, Compounds in which R ² , R ⁴ , R ⁶ and R ⁷ are hydroxyl groups and R ⁵ is a rhamnose residue (—O—C ₆ H ₁₁ O ₄ )), neoastilbin, isoastilbin and neoisoastilbin. .

The content of ampelopsin in the Fuji tea extract is usually 30% by weight or more, preferably 50% by weight or more. When the Fuji tea extract is subjected to purification treatment, the content of ampelopsin is 80% by weight or more, which is particularly preferable. In addition, the content of the taxifolin glycoside in the yellow mud extract is usually 10% by weight or more, preferably 15% by weight.
It is more than weight%. It is particularly preferable when the yellow-fruit extract is purified because the content of the taxifolin glycoside is 70% by weight or more.

Dihydroflavonols can be isolated and purified from the wisteria tea extract and / or the yellow-fruit extract, and this can be used as an active ingredient of the lipase inhibitor. The dihydroflavonols used as the active ingredient of the lipase inhibitor are not limited to those that can be isolated and purified from the wisteria tea extract and / or the yellow mulberry extract, and those isolated or purified from other plants or chemically It may be a synthetic one.

[0046]

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples.

[Production Example 1] 50 g per 200 g of dried wisteria tea branches and leaves
2000% by weight of ethanol was added, and the mixture was extracted with a reflux condenser at 80 ° C. for 1 hour and then filtered with filter paper to obtain an extract. The obtained extract was concentrated and dried under reduced pressure to obtain 50 g of Fuji tea extract (powder) (yield 25
%).

[Production Example 2] 200 g of dried wisteria tea leaves and 2 parts of water
After adding 000 ml and extracting at 90 ° C. for 1 hour,
Extraction liquid A was obtained by filtering with filter paper. Further, 2000 ml of water was added again to the extraction residue, and the mixture was similarly heat-extracted at 90 ° C. for 1 hour and then filtered through a filter paper to obtain Extract B. The obtained extracts A and B were combined to obtain an extract. The obtained extract (A + B) was passed through a glass column filled with 300 ml of a porous resin (HP-20; manufactured by Mitsubishi Chemical Corporation) to adsorb flavonoids containing amperopsin as a main component. After 600 ml of water was poured into the adsorbed porous resin to wash it, 80% ethanol was passed to elute the adsorbed component. The obtained eluate was concentrated and dried under reduced pressure to obtain 24 g (powder) of the Fuji tea extract of Production Example 2 (yield 12%). The wisteria tea extracts obtained in Production Examples 1 and 2 were subjected to high performance liquid chromatography under the following conditions to quantify the amount of ampelopsin in each wisteria tea extract.

[Conditions for high performance liquid chromatography] Detector: ultraviolet absorption detector (measurement wavelength: 290 nm) Column packing: 10 μm chemically-bonded octadecylsilane column tube: stainless steel column with an inner diameter of 4.6 mm and a length of 250 mm Temperature: 40 ° C. Mobile phase: Acetonitrile / 0.1 wt% phosphoric acid = 15/85 (weight ratio) Mixed liquid flow rate: 1 mL / min

As a result of the quantification, the amperopsin contents of the wisteria tea extracts obtained in Production Examples 1 and 2 were 38.2, respectively.
(Wt%) and 82.5 (wt%).

[Production Example 3] 70 g of dried twigs and leaves of 200 g
2000 ml of acetone by weight was added, and the mixture was extracted with a reflux condenser at 80 ° C. for 1 hour and then filtered with filter paper to obtain an extract. The obtained extract was concentrated and dried under reduced pressure to obtain 32 g (powder) of the yellow frost extract of Production Example 2 (yield 16%).

[Production Example 4] 200 g of dried twigs and leaves were mixed with 200 g of water.
After adding 000 ml and extracting at 90 ° C. for 1 hour,
Extraction liquid A was obtained by filtering with filter paper. Further, 2000 ml of water was added again to the extraction residue, and the mixture was similarly heat-extracted at 90 ° C. for 1 hour and then filtered through a filter paper to obtain Extract B. The obtained extracts A and B were combined to obtain an extract. The obtained extract (A + B) was passed through a glass column filled with 300 ml of a porous resin (HP-20; manufactured by Mitsubishi Chemical Corporation) to adsorb flavonoids containing astilbin as a main component. After 600 ml of water was poured into the adsorbed porous resin to wash it, 60% ethanol was passed to elute the adsorbed component. The obtained eluate was concentrated and dried under reduced pressure to obtain 8 g (powder) of the yellow-marble extract of Production Example 4 (yield 4
%). High-performance liquid chromatography was performed under the following conditions on the yellow-fruited extracts obtained in Production Examples 3 and 4, and the amount of astilbin in each of the wisteria tea extracts was quantified.

[Conditions for high performance liquid chromatography] Detector: ultraviolet absorption detector (measurement wavelength: 290 nm) Column packing: 10 μm chemically bonded octadecylsilane column tube: 4.6 mm inner diameter, 250 mm length stainless steel tube column Temperature: 40 ° C. Mobile phase: acetonitrile / 0.05 wt% trifluoroacetic acid = 20/80 (weight ratio) Mixed liquid flow rate: 1 mL / min

As a result of the quantification, astilbin content, neoastilbin content, neoisoastilbin content, and isoastilbin content of the yellow-fruited extract obtained in Production Example 3 were 13.1 (wt%), 1 .2 (wt%),
It was 1.0 (wt%) and 1.0 (wt%). Also,
Astilbin content of the yellow-fruited extract obtained in Production Example 4,
The content of neo-astilbin, the content of neo-iso-astilbin, and the content of iso-astilbin are 64.4 (wt%), 5.9 (wt%), 4.9 (wt%) and 4.9, respectively.
(% By weight). These results are summarized in Table 1.

[Table 1] Production Example 3 Production Example 4 Astilbin content 13.1% by weight 64.4% by weight Neoastilbin content 1.2% by weight 5.9% by weight Neoisoastilbin content 1.0% by weight % 4.9% by weight Isoastilbin content 1.0% by weight 4.9% by weight

[Examples 1 to 4] The wisteria tea extracts of Examples 1 and 2 (Examples 1 and 2) and the yellow-fruit extracts of Examples 3 and 4 (Examples 3 and 4) were mixed with 50% by weight of ethanol. Sample concentration of 1000 to 10 μg by dissolving and diluting with
/ Ml sample solution was prepared. Each sample solution 50 obtained
μl, lipase (Candida cylindracea origin, Biocataly
sts; 15.4 U / mL) 50 μl, 5,5′-dithiobis (2-nitro-benz) dissolved in 0.1 M Tris-HCl buffer (pH 8.5)
oic acid) (DTNB, 0.1 mg / mL) 340 μl and phenylmethylsulfonyl fluoride (PMS) dissolved in ethanol
F, 3.5 mg / mL) (10 μl) were mixed well and allowed to stand at 30 ° C. for 5 minutes. In this enzyme reaction solution, dissolved in ethanol 2,
3-dimercapto-1-propanol tributylate (BALB, 0.3mg / m
L) and sodium dodecylsulfate (SDS, 0.26 mg / mL) were added in an amount of 25 μl each, and then reacted at 30 ° C. for 30 minutes. After stopping the enzymatic reaction by adding 500 μl of acetone, a wavelength of 4 proportional to the amount of hydrolysis of BALB was obtained.
The absorbance at 14 nm was measured. Further, as a control, the same operation and the absorbance measurement were carried out when 50% by weight of ethanol was added instead of the sample solution. From the measurement results, the lipase activity inhibition rate was calculated by the following formula.

[0057]

[Formula 1] Lipase activity inhibition rate (%) = 1-[(A1-A
0) / (A3-A2)] × 100

In formula 1, "A0" is the absorbance when the sample solution is added (before the enzyme reaction is started), "A1" is the absorbance when the sample solution is added (after the enzyme reaction is started), and "A0" is
2 ”is the absorbance of the control (before the start of the enzyme reaction),“ A
"3" represents the absorbance of the control (after the start of the enzyme reaction).

The sample concentration of the sample solution is 6.25 to 1000.
The above-mentioned lipase activity inhibition rate was measured stepwise by changing to μg / ml, and the sample concentration (IC ₅₀ ) (μg / mL) of the sample solution at which the inhibition rate was 50% was determined. IC _{50 in} 4 is 26, 20, 2 respectively.
00 and 130 (μg / mL). The results regarding Example 1 (wisteria tea extract of Production Example 1) and Example 2 (wisteria tea extract of Production Example 2) are shown in Table 2, and Example 3 (yellow mud extract of Production Example 3) and Example. Table 3 shows the results regarding No. 4 (yellow-fruit extract of Production Example 4).

[Table 2] Sample Concentration Example 1 Example 2 (μg / mL) (Fuji Tea Extract of Production Example 1) (Fuji Tea Extract of Production Example 2) 1000 95.8 95.6 400 94. 7 91.8 100 91.3 92.6 25 47.5 56.0 6.25 19.6 0 0 0 IC ₅₀ 26 20

[Table 3] Sample Concentration Example 3 Example 4 (μg / mL) (Eye extract of Production Example 3) (Eye extract of Production Example 4) 1000 96.0 97.6 400 76. 5 94.7 100 27.6 48.2 25 8.6 16.0 6.25 4.6 0 0 0 IC ₅₀ 200 130

From the results shown in Tables 2 and 3, it was confirmed that the wisteria tea extract and the yellow eyelid extract have a strong lipase inhibitory action. In addition, it was confirmed that the wisteria tea extract having a high amperopsin content (Example 2) and the yellow musk extract having a high astilbin content (Example 4) had a stronger lipase inhibitory action.

[0063]

INDUSTRIAL APPLICABILITY The present invention provides a novel lipase inhibitor. According to the lipase inhibitor of the present invention, through the lipase inhibitory action, it is possible to prevent the decomposition of oils and fats contained in foods and drinks and cosmetics, and suppress the malodor and spoilage of foods and drinks and lipase. It is possible to prevent and treat acne and dermatitis through the inhibitory action, and prevent and treat obesity, hyperlipidemia, atherosclerosis, arteriosclerosis and the like through the inhibitory action of lipase. In addition, the lipase inhibitor of the present invention has little adverse effect on the flavor and feeling of use such as foods and drinks and cosmetics to be added, and therefore can be used for a wide range of added objects. Furthermore, since a large amount of wisteria tea and / or yellow mud, which is a raw material for extracting the active ingredient, is present in nature, and stable supply is possible,
The lipase inhibitor of the present invention can be produced inexpensively.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) // C07D 311/32 C07D 311/32 (72) Inventor Masazumi Miyakoshi 14703-10, Muto-cho, Onomichi, Hiroshima Prefecture Maruzen Pharmaceutical company F term (reference) 4C062 EE50 EE54 4C086 AA01 BA08 MA01 MA04 NA14 ZA70 ZC20 ZC33 4C088 AB12 AB56 AC05 AC06 BA08 BA09 BA32 MA07 NA14 ZA70 ZC20 ZC33

Claims (4)

[Claims] 1. A lipase inhibitor, which comprises an extract of Fuji tea and / or an extract of yellow jelly as an active ingredient. 2. The wisteria tea extract is an extract obtained by extracting the branches and leaves of wisteria tea with a polar solvent, and the yellow-branch extract is obtained by extracting the branches and leaves of yellow-brown with a polar solvent. The lipase inhibitor according to claim 1, which is the obtained extract. 3. A lipase inhibitor, which comprises dihydroflavonols as an active ingredient. 4. The lipase inhibitor according to claim 3, wherein the dihydroflavonols are ampelopsin, astilbin, neoastilbin, neoisoastilbin or isoastilbin.