JP2006241039A - Urease inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a urease inhibitor effectively inhibiting activity of urease, increased in safety to living bodies, and easily obtainable of its raw materials. <P>SOLUTION: The urease inhibitor usable in food territory, medicinal field, chemicals field and the like is disclosed. The urease inhibitor comprises, as an active ingredient, at least one selected from a group consisting of phloroglucinol, eckol, phlorofucofuroeckol A, dieckol, 8,8'-bieckol and eckstolonol. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a urease inhibitor, and more particularly to urease inhibition that catalyzes the hydrolysis of urea and effectively inhibits the activity of urease that promotes the production of ammonia.

  Urease is an enzyme that catalyzes the hydrolysis of urea and produces ammonia. Such activity of urease is said to cause problems in various aspects.

  One problem is gastritis. Helicobacter pylori (H. pylori), which survives in the stomach, decomposes urea into ammonia by its own urease, resulting in H. pylori. Pylori-positive people have a significantly higher ammonia concentration in gastric juice than negative people, and there is a risk that the gastric mucosa may be damaged by this. In actual clinical practice, it has been reported that there is a significant correlation between the ammonia concentration in gastric juice and the gastritis score. In vitro ammonia damage to gastric epithelial cells has also been demonstrated. Therefore, H.H. As a therapeutic agent for gastric mucosal injury due to long-term ammonia exposure predicted in patients with pylori-positive gastritis or gastric ulcers, development of urease inhibitors has been considered (for example, see Non-patent Document 1).

  As another problem, hepatic coma based on cirrhosis has been pointed out. Hepatic coma is a disturbance of consciousness observed at the time of liver disorders such as cirrhosis, and the main factor is thought to be ammonia. About 70% of the urea synthesized in the liver is excreted as it is in the urine via the kidney, and the remaining about 30% of urea is secreted into the digestive tract. When urea secreted in the digestive tract is decomposed by urease present in the digestive tract, ammonia is produced. Thereafter, the generated ammonia is absorbed into the living body, and most of it is re-synthesized into urea in the liver. However, in patients suffering from liver diseases such as cirrhosis, the ability to synthesize urea in the liver is reduced, which may lead to an increase in blood ammonia concentration and development of consciousness disorder (hepatic coma). Yes. In this way, the cause of hepatic coma is ammonia, and gastrointestinal urease has been pointed out as one of the causes of increased blood ammonia concentration. Therefore, it is thought that inhibition of urease is effective in treating hepatic coma (For example, refer to Patent Document 1).

  As another problem, diaper smell and diaper rash have been pointed out. Urease present in stool breaks down urea in urine and produces ammonia. When feces and urine are mixed in a diaper, ammonia is produced by the reaction of urease and urea, which diffuses into the atmosphere and gives off a bad odor. It is also known that the ammonia produced increases the skin pH and activates fecal proteases and lipases to induce diaper rash. In order to suppress or prevent such ammonia odor and diaper rash, Patent Document 2 discloses an ammonia odor inhibitor and a diaper rash prevention containing compounds such as flavones, flavonols, dihydroflavonols, and catechins as active ingredients. An agent is disclosed.

  While various problems have been pointed out as described above, various urease inhibitors have been proposed and developed for the purpose of inhibiting the activity of urease. As typical urease inhibitors, for example, those using a benzimidazole compound (Patent Document 3), those using a carbostyl derivative (Patent Document 4), those using a dithiodibenzohydroxamic acid derivative (Patent Document 5), Those using thiazole derivatives (Patent Document 6), those using plant extracts such as Lucas (Patent Document 7), those using plant extracts such as Diaus (Patent Document 8), and plant extracts such as Acacia (Patent Document 9).

  However, urease inhibitors using the above components are not necessarily safe in terms of the presence or absence of side effects on the living body, and urease inhibitors using components derived from plant extracts, in industrial production, Considering whether it is relatively easy to obtain the plant itself as a raw material and whether it has an effective inhibitory effect on the activity of urease, it is still sufficient in this respect. hard.

Takahashi et al., “Ulcer Research”, 2003, Vol. 106-109 JP-A-6-256375 JP 2004-091338 A Japanese Unexamined Patent Publication No. 7-025767 JP-A-7-101862 Japanese Patent Laid-Open No. 10-316651 JP 2001-247462 A Japanese Patent Laid-Open No. 8-019595 JP 2000-154145 A JP 2002-029978 A

  An object of the present invention is to solve the above-mentioned problems. The purpose of the present invention is to more effectively inhibit the activity of urease, improve the safety to the living body, and obtain the raw materials. It is an object to provide a urease inhibitor that is easy.

  The present invention includes urease containing, as an active ingredient, at least one phenol compound selected from the group consisting of phloroglucinol, equol, florofcofurecol A, diecol, 8,8'-biequol, and extronol. An inhibitor.

  In one embodiment, the phenol compound is derived from seaweed.

  In one embodiment, the phenolic compound is contained in the form of a seaweed extract.

  In a further embodiment, the seaweed is a brown seaweed albinocephalum macaque, a brown algae albinoceae arame, sagarame, tsuruarame, kurome, kajime, wakame, aowakame, hirome, ainu wakame, and chigaiso; At least 1 selected from the group consisting of Hidawara, Hijiki, and Akamok of the Hiba or Hidawara family; Mozuku and Okinawa Mozuku of the brown algae moths and scabies; and a seaweed belonging to the red alga Sugiori It is a species of seaweed.

  The present invention is also a food or drink with a label indicating that it is a food for inhibiting the activity of urease, phloroglucinol, equol, florofcofurecole A, diecol, 8,8′-biequol, and A food or drink containing as an active ingredient at least one phenol compound selected from the group consisting of extronol.

  In one embodiment, the phenol compound is derived from seaweed.

  According to the present invention, the activity of urease, which catalyzes the hydrolysis of urea to produce ammonia, can be more effectively inhibited, and the safety to the living body can be improved. For this reason, the urease inhibitor of the present invention can be more routinely used, and versatility can be enhanced in various fields such as the food field and the pharmaceutical field. The urease inhibitor of the present invention is also used to prevent or treat various diseases such as gastritis, hepatic coma based on cirrhosis, diaper odor, diaper rash, etc. that can be caused by urease catalyzing and decomposing urea into ammonia. Can be used for purposes.

  First, the urease inhibitor of the present invention will be described.

  The urease inhibitor of the present invention contains a phenol compound as an active ingredient.

  The phenol compound used in the present invention is a kind of polyphenol which is also called phloroglucinol itself or seaweed tannin (phlorotannin). Furthermore, this polyphenol is a compound that includes the entire fluorotannins having phloroglucinol as a basic skeleton unit. As a phenol compound used for this invention, the following formula | equation:

Phloroglucinol itself, equol, florofucofurequol A, diecol, 8,8'-biequol, exotronol Fluorotannins exemplified in (eckstronol), and combinations thereof.

  The phenolic compound used in the present invention is commercially available, or can be easily obtained from seaweed as described later using techniques well known to those skilled in the art. In particular, the phenolic compound used in the present invention is preferably derived from seaweed because it is easily available. Moreover, the phenol compound used for this invention may be used with the form of a seaweed extract.

  Examples of seaweeds that can be used in the present invention include the brown algae Kombu Kombu family, the brown alga Kombu Komiso family, the brown algae Hiba or Homidae family, the brown algae Nagamatsu and Mozaku family, or the red alga Suginori. Examples include seaweeds that belong to the department of super eyes.

  Specific examples included in the seaweed include the brown algae Kombu Kombu (Laminaria japonica), the brown algae Osmophila Alame (Eisenia bicyclis), Sagarame (Eisenia arborea), tsuru arame stlonifera), chromate (Ecklonia kurome), beforehand (Ecklonia cava), seaweed (Undaria pinnatifida), Aowakame (Undaria peterseniana), spread (Undaria undarioides), Ainu wakame (Alaria praelomga), and Chigaiso (Alaria crassifolia); brown algae hiba In addition, Honda Wara (Sar gassium fluvellum, hijiki fusiforme, and red moss (Sargassum horneri); brown algae (Nemacytus decipieus) and red moss Seaweed; In particular, because the production of seaweed itself is relatively large and easily available, and when the phenolic compound is used in the form of a seaweed extract, it is easier to obtain a fraction containing more of the phenolic compound. Alame, chrome, wakame and hijiki are preferred.

  In the present specification, the seaweed includes the whole grass of these seaweeds themselves or at least one part including roots, stems, and leaves. The seaweed forms used in the present invention include raw ones, fragments, strips or powders. Furthermore, the seaweed that can be used in the present invention includes both dry matter and raw state.

  Moreover, in this specification, an extract means the extract extracted on suitable conditions using the above seaweeds as water, a polar or nonpolar solvent, or these mixtures as an extraction solvent. The form of the extract is not particularly limited, and includes an extract, or a powder or paste obtained by concentrating or drying the extract by means usually used by those skilled in the art.

  In the present invention, when the phenol compound is used by being taken out from an extract derived from seaweed or used in the form of the extract, the seaweed-derived extract is preferably (1) by extracting the seaweed with water. Or (2) 10 (v / v) of the seaweed, which is derived from the supernatant of the C1-C3 alcohol liquid containing 50 (v / v)% or more of the obtained crude water extract. % Of C1-C3 alcohol or alcohol extract obtained by extraction in an aqueous alcohol solution.

  The above (1) “derived from the supernatant of a C1 to C3 alcohol solution of 50 (v / v)% or more containing a crude water extract obtained by extracting seaweed with water” that can be used in the present invention. Is obtained by extracting the above-mentioned seaweed once in water through an operation described below to obtain a crude water extract, and then combining this with a C1-C3 alcohol or a C1-C3 alcohol aqueous solution, 50% (v / v) It can be obtained by fractionating the supernatant from a C1-C3 alcohol solution of% or more. Examples of C1-C3 alcohols include methanol, ethanol, and propanol.

  On the other hand, the above (2) “derived from an alcohol extract obtained by extracting 10% (v / v)% or more of a C1-C3 alcohol or an aqueous alcohol solution”, which can be used in the present invention, The seaweed is treated with C1-C3 alcohol itself or a predetermined concentration (preferably 10 (v / v)% to 80 (v / v)%, more preferably 15 (v / v)% to 50 ( It can be obtained by extraction in a C1-C3 alcohol aqueous solution adjusted to v / v)%). Examples of C1-C3 alcohols include methanol, ethanol, and propanol.

  The seaweed-derived extract is preferably 10% by weight or more, more preferably 15% by weight to 100% by weight, and still more preferably 30% by weight, based on the weight of the solid content contained in the extract. % To 99% by weight, still more preferably 60% to 99% by weight. Here, the term “weight of solids contained in the extract” used in the present specification means that the liquid component contained in the extract is removed from the extract (for example, evaporated) to obtain a solid. Refers to the weight of a substance that can remain as a component. When the extract satisfies the phenol compound content in such a range, the inhibitory action on urease is remarkably improved.

  In the urease inhibitor of the present invention, the seaweed-derived extract containing more of the phenol compound used in the inhibitor is not particularly limited. For example, the following first method or second method is used. Can be manufactured.

  The first method will be described.

  In the first method, first, a water crude extract is produced by immersing the seaweed in water for a predetermined time.

  Although the quantity ratio (extraction capacity) of the seaweed and water in this immersion is not particularly limited, for example, it is preferably 1 liter to 10 liters, more preferably 2 liters to 5 liters with respect to 100 g of seaweed (dry weight). Water is used.

  The water used is, for example, room temperature water (particularly neither heated nor cooled) or hot water, and particularly when hot water is used, the temperature (extraction temperature) is preferably 60 ° C. to 100 ° C., More preferably, it is 80 degreeC-100 degreeC, More preferably, it is 90 degreeC-100. In addition, while the seaweed is immersed, for the purpose of facilitating the extraction of the phenol compound used in the present invention, means known to those skilled in the art are used so as not to lower the temperature of hot water. It is preferable to maintain the temperature by heating.

  The time of immersion in water (extraction time) varies depending on the extraction temperature to be used, and is not necessarily limited. For example, when hot water is maintained at about 100 ° C., preferably 1 minute to 120 minutes, more preferably 10 Min to 60 min. By carrying out the extraction time within such a range, the phenol compound used in the present invention can be extracted more efficiently, and excessive extraction of unnecessary substances can be prevented.

  After the immersion, for example, it is allowed to cool to room temperature, and seaweed is removed by filtration or centrifugation. Thus, a crude water extract can be obtained. The obtained crude water extract may be derived from an aqueous layer from which the organic layer has been removed by combining with an organic solvent such as hexane or chloroform for the purpose of removing impurities in advance. Furthermore, the obtained water crude extract may be used as it is for the steps described below, or, if necessary, a dried or pasty product obtained by evaporating water using means known to those skilled in the art. It may be used in the form of

  Next, the water crude extract is combined with the C1-C3 alcohol or the C1-C3 alcohol aqueous solution, preferably 50 (v / v)% or more, more preferably 60 (v / v)% to 98 (v / v). C1-C3 alcohol liquid having the alcohol concentration of%) is prepared. Here, the concentration of the C1-C3 alcohol solution that can be prepared can be set to any concentration by those skilled in the art, preferably within such a range. That is, it is conceivable that the content of the phenolic compound that inhibits urease, which is useful in the present invention, varies depending on the type of seaweed used, the site of use, the production area, the collection time, the storage state after collection, and the like. In addition, an error may occur in the obtained component content even in a series of extraction operations described later. Therefore, a person skilled in the art can arbitrarily set the concentration of the alcohol liquid that can be prepared within such a range in accordance with such conditions.

  More specifically, the water crude extract is combined with an alcohol having 1 to 3 carbon atoms or an aqueous alcohol solution having 1 to 3 carbon atoms (hydrous alcohol) prepared at a predetermined concentration. Thus, an alcohol solution having the alcohol concentration as described above is prepared. Examples of alcohols having 1 to 3 carbon atoms that can be used in such operations include methanol, ethanol and propanol, and combinations thereof. In consideration of further improving the safety to the living body, it is preferable to use ethanol. The amount of such alcohol or hydrous alcohol used is not particularly limited because it varies depending on the amount of the crude water extract to be combined.

  Thereafter, the supernatant of the C1-C3 alcohol solution is collected.

  By preparing an alcohol solution set to the alcohol concentration, a substance insoluble in the alcohol solution may precipitate. The supernatant is collected for the purpose of removing this precipitate, and the supernatant can be removed by methods well known to those skilled in the art (for example, filtration or centrifugation).

  Next, the second method will be described.

  In the said 2nd method, extraction is performed by immersing the said seaweed in C1-C3 alcohol or alcohol aqueous solution of 10 (v / v)% or more for a predetermined time, and an alcohol extract is manufactured.

  The C1-C3 alcohol or alcohol aqueous solution used above is preferably 10 (v / v)% to 80 (v / v)%, more preferably 15 (v / v)% to 50 (v / v)%. It is preferable that it is C1-C3 alcohol aqueous solution set to.

  The quantity ratio (extraction capacity) between the seaweed and hot water in this immersion is not particularly limited, but is preferably 0.5 liters to 5 liters, more preferably 0.5 liters with respect to 100 g of seaweed (dry weight), for example. 1 to 2 liters, even more preferably 1 liter of the above C1 to C3 alcohol or aqueous alcohol solution is used.

  Although the temperature (extraction temperature) of the alcohol used is not specifically limited, Preferably it is 0 degreeC-50 degreeC, More preferably, it is 1 degreeC-30 degreeC, More preferably, it is 15 degreeC-25 degreeC.

  The time (extraction time) for immersing in the C1-C3 alcohol or alcohol aqueous solution is preferably 2 hours to 48 hours, more preferably 4 hours to 24 hours. By carrying out the extraction time within such a range, the phenol compound used in the present invention can be extracted more efficiently, and excessive extraction of unnecessary substances can be prevented.

  After the immersion, seaweed is removed by centrifugation or filtration. In this way, an alcohol extract can be obtained. The obtained alcohol extract may be derived from an aqueous layer from which the organic layer has been removed by combining with an organic solvent such as hexane or chloroform for the purpose of removing impurities in advance. Further, the obtained alcohol extract may be used as it is for the purification step described later, or, if necessary, in the form of a concentrate obtained by evaporating alcohol or water using means known to those skilled in the art. May be used.

  In this manner, an extract derived from seaweed containing more of the phenol compound used in the present invention can be produced by using either the first method or the second method. The obtained seaweed-derived extract may be purified for the purpose of enhancing the urease inhibitory action in the present invention. This purification is performed by, for example, concentrating the supernatant obtained above using a means well known to those skilled in the art, and then preferably 40 (v / v)% to 90 (v / v) )%, More preferably 50 (v / v)% to 80 (v / v)% C1-C3 alcohol (preferably ethanol) aqueous solution is passed through a column used as an eluent. The adsorbent useful for this column chromatography is preferably an aromatic adsorbent, and a more specific example is a styrene-divinylbenzene adsorbent. The styrene-divinylbenzene adsorbent is commercially available from Mitsubishi Chemical Corporation under the trade name Diaion HP20, for example.

  By performing the chromatography, it is possible to extract a fraction having a more enhanced urease inhibitory effect (that is, the content of the phenol compound used in the present invention is increased). The obtained fraction can be used as it is as the urease inhibitor of the present invention.

  In addition, when further isolating the phenol compound used in the present invention from the seaweed extract or fraction obtained above, for example, the following second column chromatography (for example, T. Nakamura et al., Fishers) Science (see Sciences, 1996, Vol. 62, p. 923-926).

  That is, the seaweed extract or fraction obtained above is applied to a column packed with, for example, silica gel as an adsorbent. Examples of eluents used include chloroform / methanol / distilled water combinations. Furthermore, the obtained second fraction is obtained by, for example, thin-layer chromatography using silica gel as a thin layer plate and chloroform / methanol / distilled water / acetic acid as a developing solvent, and the above phloroglucinol, equol, fluoro Fucophoreol A, diecol, 8,8′-biequol, and exonol can be isolated respectively.

  The urease inhibitor of the present invention can be used for any purpose, either for oral administration or ingestion, or for external skin application. Further, the urease inhibitor of the present invention is, for example, an additive for deodorizing or deodorizing and / or an additive for preventing rash or rough skin in hygiene products (for example, paper diapers, sanitary products, wet tissues). Can be used as The urease inhibitor of the present invention is not limited only to such sanitary products, but prevents or reduces the occurrence of unpleasant odor (ammonia odor) caused by the occurrence or occurrence thereof in a place where ammonia is generated from urea by the action of urease, Alternatively, it can be used for suppressing or reducing skin damage caused by contact with the skin.

  The urease inhibitor of the present invention may also contain other components depending on the purpose in addition to the seaweed-derived extract as an active ingredient. Examples of other ingredients that may be included in the present invention include water; alcohol; processed meat products; rice, wheat, corn, potatoes, sweet potatoes, soy, kombu, wakame, tengusa and other common food materials and their powders; Sugars such as starch, starch syrup, lactose, glucose, fructose, sucrose, mannitol; general food additives such as spices, sweeteners, edible oils, vitamins; surfactants; excipients; coloring agents; preservatives; Coating aids; lactose; dextrin; corn starch; sorbitol; crystalline cellulose; polyvinyl pyrrolidone; oils; moisturizers; thickeners; The urease inhibitor of the present invention may further contain other drugs (including traditional Chinese medicine) as necessary. The content of such other components and / or other drugs is not particularly limited, and an appropriate amount can be selected by those skilled in the art.

  Furthermore, the urease inhibitors of the present invention are not necessarily limited to either in vivo or in vitro, and can be widely used in any application where it is desired to further inhibit the activity against urease. That is, the urease inhibitor of the present invention is not particularly limited, but may be used, for example, as a kind of additive added to food compositions such as health foods, and is used in production fields such as livestock or farmed fish. The feed composition may be used as it is or in combination with other feed materials, or may be used as it is or in combination with other pharmaceutical compositions as a pharmaceutical composition such as pharmaceuticals and quasi drugs. In addition, it may be used as a kind of additive added to a base material (chemical product base material) constituting a sanitary product such as a diaper, a wet tissue, and a sanitary product.

  When the urease inhibitor of the present invention is used as a food composition, the form is not limited to a fixed food, but includes a beverage (for example, a liquid beverage). More specific examples include tea drinks, coffee drinks, soft drinks, milk drinks, confectionery, syrups, processed fruit products, processed vegetable products, pickles, and livestock meat in the form of liquids, pastes, solids, etc. Examples include, but are not limited to, products, fish products, delicacies, cans / bottles, instant food / drinks, oral liquids, liver oil drops, mouth fresheners, and jelly. Such food compositions containing the urease inhibitor of the present invention can be produced using techniques known to those skilled in the art.

  When the urease inhibitor of the present invention is used as an external preparation, its form is not particularly limited and can be used in the form of liquid, gel, powder, etc., and the inhibitor can be a lotion, It can also be blended in sprays, mousses and the like.

  When the urease inhibitor of the present invention is used for a chemical substrate, the usable substrate is not particularly limited, and examples thereof include fabrics such as woven fabrics, knitted fabrics, and nonwoven fabrics; papers;

  In addition, when the urease inhibitor of the present invention, or the above-mentioned phenol compound or a seaweed-derived extract containing the above-mentioned phenol compound is contained in the food composition, the food composition displays an indication that it has various functions. It can be used as attached food (for example, food for specified insurance or food for special use). Examples of functions attached to such indications include, but are not limited to, the expression method itself; inhibit urease activity; prevent gastritis; prevent, eliminate or alleviate stomach discomfort; Foods suitable for those who are concerned about stomach condition such as stomach pain and stomach discomfort; Prevent or treat hepatic coma; The display only needs to be expressed in a manner that allows the user to substantially understand the functions as described above. For example, the display can be performed on the exterior or interior package of the food, a product catalog, a poster, or the like.

  When the urease inhibitor of the present invention is used as a pharmaceutical composition, its dosage form is not particularly limited, and is processed into an appropriate dosage form according to the method described in the Japanese Pharmacopoeia. More specific examples of dosage forms include, for pharmaceutical compositions intended for oral administration, dosage forms such as capsules, tablets, powders, granules, fine granules, sustained release agents, and liquids. In the case of pharmaceutical compositions intended for parenteral administration, dosage forms such as injections, infusions, eye drops, ointments, creams, patches and the like can be mentioned. Since the dose can be easily varied depending on conditions such as the body weight of the subject person, it can be appropriately selected by those skilled in the art.

  The urease inhibitor of the present invention can be produced by a method usually used in the art depending on the form of use, and can be appropriately ingested or applied by a method according to the form. In this field, urease includes urease derived from peanut, H. coli. It is known that various types such as urease derived from Pylori and various ureases derived from intestinal bacteria exist. In addition, it is known that these ureases have some difference in affinity for urea as a substrate depending on their origin, but the urease inhibitor of the present invention depends on the type of urease and / or its origin. Therefore, the action of any urease can be effectively inhibited.

  Hereinafter, the present invention will be described specifically by way of examples. However, the present invention is not limited by these.

<Example 1: Production of a chrome extract containing a phenol compound>
400 g of dried chrome powder was added to 1200 mL of methanol and extracted at room temperature for 24 hours. Subsequently, the crude extract was separated by centrifugation (3000 rpm, 20 minutes), concentrated on a rotary evaporator, and dried. Thereafter, 240 mL of methanol, 480 mL of chloroform and 180 mL of distilled water were added to the dried product thus obtained, and the mixture was separated to take out the upper layer. Further, 300 mL of ethyl acetate was added to the extracted upper layer, followed by liquid separation, and the ethyl acetate layer was extracted. This separation with ethyl acetate was repeated twice. The combined ethyl acetate layer was concentrated with a rotary evaporator and dried to obtain a chrome extract (k1) containing a phenol compound (yield: 3.2% based on the weight of the dry powder used). .

<Example 2: Isolation of phenol compound (fluorotannin) from chrome extract>
10 g of the chrome extract (k1) obtained in Example 1 Nakamura et al., Fishers Science, 1996, 62, p. In accordance with the method described in 923-926, it was added to a column (10 mmφ × 300 mm) containing Wakogel C-300 (manufactured by Wako Pure Chemical Industries, Ltd.), and chloroform / methanol / distilled water (volume ratio 50) as an eluent. : 30: 7).

Trimeric equol, which is a component of the phenol compound used in the present invention, could be separated by the column chromatography. For other phenolic compounds, the fraction obtained by the column chromatography was subjected to thin layer chromatography (TLC), the corresponding spots were scraped, and eluted and purified with methanol, respectively. Fluorocofloecole A, hexamer diecol, and hexamer 8,8′-biequol were separated. The conditions used for this TLC are as follows: Silica gel 60F ₂₅₄ (Merck) is used as the thin layer plate, and chloroform / methanol / distilled water / acetic acid (volume ratio 50: 25: 4) as the developing solvent. : 3) and detected at UV 254 nm.

  The yields of the obtained equol, Fluorocofurol col A, diecol, and 8,8'-bicol are 6%, 7%, 10% and 8%, respectively, based on the weight of the chrome extract (k1) used. 2%.

<Example 3: Evaluation of urease inhibitory activity>
The various phenol compounds obtained above were dissolved in 50% ethanol and diluted sequentially in two stages using distilled water to obtain diluted sample solutions of the various phenol compounds.

  10 μl of the sample solution is placed in a microplate, and 10 μl of 47.5 U / ml juvenile urease (manufactured by Wako Pure Chemical Industries, Ltd.) is added to the microplate and preincubated at 37 ° C. for 10 minutes. did. Next, the reaction solution containing urea (107 mg of urea dissolved in 10 mL of phosphate buffer (pH 7.0), 2 g of sodium salicylate and 0.1 g of sodium nitroprusside was added to 50 ml of phosphate buffer (pH 7). 0.0) with a volume ratio of 1: 7) and 80 μl were added and incubated at 37 ° C. for 30 minutes. Thereafter, 80 μl of an ammonia reaction reagent (a solution prepared by dissolving 50 μl of hypochlorous acid and 0.75 g of sodium hydroxide in 50 ml of phosphate buffer (pH 7.0)) was added to the microplate, After 10 minutes of incubation at 0 ° C., the absorbance at 570 nm of the obtained evaluation sample solution was measured.

  On the other hand, as a control, a control evaluation was performed in the same manner as in the above evaluation method except that the various phenol compounds were not added. Furthermore, blank evaluation was performed similarly to the said evaluation method except having used 10 microliters phosphate buffer (pH 7.0) instead of the urease solution as a blank.

  For the absorbance of the evaluation sample solution, control evaluation solution, and blank evaluation solution obtained above, the following formula:

Inhibition of urease of phenol compounds of equol, Fluorocofloecole A, diecole, and 8,8′-biequol obtained in Example 2 above, and phloroglucinol (manufactured by Wako Pure Chemical Industries, Ltd.) The rate (%) was calculated, and the concentration (IC ₅₀ ) of the various phenol compounds that inhibited the used urease by 50% was determined from the relationship between the concentration of the various phenol compounds used and the inhibition rate. The obtained results are shown in Table 1. The IC ₅₀ value of the chrome extract (k1) was 6.4 μg / mL.

  Furthermore, 10 mg of the chrome extract (k1) obtained above was accurately weighed, and the total phlorotannin amount of the chrome extract was determined by the Folin-Denis method using phloroglucinol as a standard substance (Journal of Japanese Society of Food Science and Technology) , 2002, vol. 49, pp. 507-511). The total amount of fluorotannin in the chrome extract was 115.2%.

  As shown in Table 1, all of equol, fluorocofloecole A, diecole, 8,8′-biequol, and phloroglucinol included in the phenol compound used in the present invention are excellent in microgram order. It can be seen that it has urease inhibitory activity. Furthermore, equol, florofoflor equol A, diecol, and 8,8′-biequol contained in the chrome extract obtained in Example 1 have particularly superior urease inhibitory activity over commercially available phloroglucinol. It has been found that the chrome extract containing these fluorotannins (phenolic compounds) is also useful as an excellent urease inhibitor.

<Example 4: Preparation of food composition>
A food composition having the following composition was prepared using the chrome extract obtained in Example 1 above.

Ingredient Weight (g)
Chromome extract obtained in Example 1 0.6
Soy Saponin 2.0
Black vinegar extract 2.0
Apple fiber 2.0
Lecithin 1.0
Fructooligosaccharide 2.0
Fructose 1.0
Powdered vinegar 0.1
Cyclodextrin 1.0
Honey 1.0
Bone meal 1.0
Dextrin 4.9.

  Each component was mixed in a fluid granulator, granulated by spraying water, and dried at an inlet temperature of 80 ° C. to obtain a granular food product.

  The urease inhibitor of the present invention contains a phenolic compound contained in seaweed, which can be widely used as food, as an active ingredient, so it is highly safe and can be used on a daily basis. The versatility in various fields, such as a field, can be improved. The urease inhibitor of the present invention also prevents or treats various diseases that can be caused by urease catalyzing and decomposing urea into ammonia, such as gastritis, hepatic coma based on cirrhosis, diaper odor, diaper rash, etc. It can be used for the purpose of eliminating discomfort due to deodorization of the ammonia odor.

Claims (6)

  A urease inhibitor comprising, as an active ingredient, at least one phenol compound selected from the group consisting of phloroglucinol, equol, florofcofurecol A, diecol, 8,8'-biequol, and extronol.   The urease inhibitor according to claim 1, wherein the phenol compound is derived from seaweed.   The urease inhibitor according to claim 1, wherein the phenol compound is contained in the form of a seaweed extract.   The seaweeds are brown algae (Homoptera), Kombu (Brassica), brown algae (Homoptera), Arame, Sagarame, Tsuruarame, Kurome, Kajime, Wakame, Aowakame, Hirome, Ainu Wakame, and Chigaiso; Brown algae At least one kind of seaweed selected from the group consisting of: Alaska honda, hijiki, and akamoku; brown algae, moss and oak, and red seaweeds The urease inhibitor according to claim 2 or 3.   A food or drink with a label indicating that it is a food for inhibiting the activity of urease, comprising phloroglucinol, equol, florofcofurecole A, diecole, 8,8'-biequol, and extronol A food or drink comprising as an active ingredient at least one phenol compound selected from the group.   The food and drink according to claim 5, wherein the phenol compound is derived from seaweed.