JP6211406B2 - Muscarinic receptor activator and salivary secretion promoter - Google Patents

Description

  The present invention relates to a muscarinic receptor activator and a salivary secretion promoter.

  Asparathus linearis is also called rooibos, belongs to the genus Fabaceae, Aspalathus, and has coniferous leaves. Tea made by drying the leaves is known as a kind of healthy tea called "Rooibos tea".

  Patent Document 1 discloses that rooibos tea is effective in alleviating insomnia, nervous tension, gastric spasm, and allergic symptoms. Flavonoids in rooibos tea have strong antioxidant activity and free radicals. It is described that it has scavenge activity and is known to have the potential to act as an anticancer agent and an anti-atherosclerotic agent. Patent Document 1 describes that rooibos tea is useful as an antidiabetic agent.

  Patent Document 2 describes an adiponectin production inhibitor containing an extract of rooibos leaf, and describes that the adiponectin production inhibitor is used for slimming applications.

  Patent Document 3 describes an oral composition containing an Asparagus lineus extract as a bad breath deodorant, and describes the use of the composition as an oral wetting agent.

  Patent Document 4 describes an oral antibacterial agent containing luteolin as an active ingredient.

  Dry mice are also experienced in daily life, accompanied by sticky discomfort, difficulty in conversation, and the generation of bad breath. Further morbidity results in not only oral function but also general health failure such as caries, periodontal disease, various infections, etc. due to changes in oral flora. Therefore, promoting the secretion of saliva to moisturize the oral cavity is important for keeping the oral cavity refreshing and preventing oral and systemic diseases.

  Patent Document 5 describes that polyglutamic acid and its salts exhibit excellent salivary secretion promoting activity and moisturizing action. However, polyglutamic acid and its salt have a high viscosity, which may cause a problem when formulated.

Special table 2010-520913 JP 2008-24620 A JP 2006-117563 A JP 2000-239136 A International Publication No. 2005/049050

  Therefore, since a search for a substance effective for reducing dry mouth has been sought, the present invention has an object to provide a substance effective for reducing dry mouse.

  The inventors of the present invention have made extensive studies in view of the above problems. As a result, the extract of Asparathus linearis and luteolin activates a muscarinic receptor present in salivary gland cells. It has been found that the secretion is promoted, and the present invention has been completed.

The present invention provides the following inventions.
[1] A muscarinic receptor activator, comprising at least one member selected from the group consisting of an extract of Asparathus linearis and luteolin.
[2] A salivary secretion promoter comprising at least one selected from the group consisting of an extract of Asparathus linearis and luteolin.
[3] A composition for activating muscarinic receptors or promoting salivary secretion, comprising the agent according to [1] or [2] as an active ingredient.
[4] The composition according to [3], comprising 1 to 50% by mass of at least one member selected from the group consisting of an extract of Asparathus linearis and luteolin.
[5] From a group consisting of a muscarinic receptor activating action and a salivary secretion promoting action in a food, comprising adding at least one selected from the group consisting of an extract of Asparathus linearis and luteolin to the food A method of imparting at least one selected action.

  The Asparathus linearis extract and luteolin according to the present invention have a muscarinic receptor activation action and a salivary secretion promoting action, and can effectively ameliorate the symptoms of dry mice.

FIG. 1 is a graph showing the activation effect of luteolin on muscarinic receptors (Example 1). FIG. 2A is a graph showing the effect of apiin on muscarinic receptors (Example 1). FIG. 2B is a graph showing the action of aspartin on muscarinic receptors (Example 1). FIG. 2C is a graph showing the effect of apigenin on muscarinic receptors (Example 1). FIG. 2D is a graph showing the effect of quercetin on muscarinic receptors (Example 1). FIG. 2E is a graph showing the effect of luteolin-7-O-glucoside on muscarinic receptors (Example 1). FIG. 2F is a graph showing the effect of luteolin-8-C-glucoside on muscarinic receptors (Example 1). FIG. 2G is a graph showing the effect of 4 ', 6,7-trimethoxyisoflavone on muscarinic receptors (Example 1). FIG. 2H is a graph showing the effect of 3 ', 4', 7-trimethoxyflavone on muscarinic receptors (Example 1). FIG. 2I is a graph showing the effect of daidzin on muscarinic receptors (Example 1). FIG. 2J is a graph showing the effect of daidzein on muscarinic receptors (Example 1). FIG. 2K is a graph showing the action of genistein on muscarinic receptors (Example 1). FIG. 2L is a graph showing the effect of genistin on muscarinic receptors (Example 1). FIG. 2M is a graph showing the effect of glycitin on muscarinic receptors (Example 1). FIG. 2N is a graph showing the effect of glycitein on muscarinic receptors (Example 1). FIG. 2O is a graph showing the effect of ipriflavone on muscarinic receptors (Example 1). FIG. 2P is a graph showing the effect of 5,7,4'-trimethoxyflavone on muscarinic receptors (Example 1). FIG. 3 is a graph showing the salivary secretion promoting effect (mouth rinse) of rooibos tea extract (Example 2). FIG. 4 is a graph showing the salivary secretion promoting effect (internal use) of rooibos tea extract (Example 3). FIG. 5 is a graph showing the actual feeling (internal use) of saliva secretion by rooibos tea extract (Example 3).

In one embodiment of the present invention, there is provided a muscarinic receptor activator comprising at least one member selected from the group consisting of an Asparathus linearis extract and luteolin.
In another embodiment of the present invention, there is provided a salivary secretion promoter characterized in that it contains at least one member selected from the group consisting of Asparathus linearis extract and luteolin.

<Asparathus linearis extract>
Asparathus linearis is also called rooibos (hereinafter simply referred to as “rooibos”) and belongs to the genus Fabaceae asparatus.

  The extraction site of rooibos is not particularly limited and may be appropriately selected as long as the desired effect of the present invention can be achieved. For example, leaf, young leaf, branch, trunk, bark, flower , Fruit part, root part and the like. Among these, a young leaf part and a branch part are preferable.

  As a method for preparing the rooibos extraction site, it can be obtained by fermentation treatment, pulverization using a crusher, or drying treatment, or a combination thereof, followed by solvent extraction described later. The fermentation treatment is not particularly limited, and a generally used method can be used. For example, it is preferable to incubate at 20 to 40 ° C. for 1 to 24 hours after water is added. The drying process is not particularly limited, and a generally used method can be used. For example, it is preferable to carry out at 60 degrees C or less.

  The rooibos extract can be easily obtained by a method generally used for plant extraction. The specific embodiment of the rooibos extract is not particularly limited and may be appropriately selected as long as the desired effect of the present invention can be achieved. For example, the extract itself, the dried extract, and the extract dilution Liquid, dried extract dilution, concentrated extract (concentrated extract), dried extract concentrate, and the like can be used.

  The method for extracting rooibos is not particularly limited and may be appropriately selected as long as the desired effect of the present invention can be achieved. For example, the extraction part of rooibos as an extraction raw material is put into a processing tank filled with an extraction solvent, and after eluting soluble components while stirring as necessary, the rooibos extract is removed by filtration to remove the extraction residue Can be obtained.

  The extraction conditions (extraction time and extraction temperature) of rooibos and the amount of extraction solvent used for rooibos are not particularly limited and are appropriately selected depending on the extraction method and the like as long as the desired effect of the present invention can be achieved. be able to.

  The extraction solvent for rooibos is not particularly limited and may be appropriately selected as long as the desired effect of the present invention can be achieved. For example, water, a hydrophilic solvent, a hydrophobic solvent, or a mixed solvent thereof may be used. Can be mentioned. There is no restriction | limiting in particular as water, According to an administration route, an administration method, etc., it can select suitably. For example, pure water, tap water, well water, mineral water, mineral water, hot spring water, spring water, fresh water, purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, etc. Can be mentioned. There is no restriction | limiting in particular as a hydrophilic solvent, According to an administration route, an administration method, etc., it can select suitably, For example, C1-C6 lower alcohols, such as methanol, ethanol, propyl alcohol, isopropyl alcohol; Examples include lower aliphatic ketones such as acetone and methyl ethyl ketone; polyhydric alcohols having 2 to 6 carbon atoms such as 1,3-butylene glycol, propylene glycol, and glycerin. Examples of the hydrophobic solvent include aromatic carbon such as benzene and toluene; organic solvent such as ethyl acetate, acetonitrile and ether; halogenated carbon such as dichloromethane and chloroform. There is no restriction | limiting in particular as a mixed solvent, The said water, the said hydrophilic solvent, etc. can be mixed suitably and used. When lower alcohol is used, 1 part by weight to 90 parts by weight with respect to 10 parts by weight of water, and when lower aliphatic ketone is used, 1 part by weight to 90 parts by weight with respect to 10 parts by weight of water, polyvalent When using alcohol, it is preferable to add 1-90 mass parts with respect to 10 mass parts of water. The rooibos extraction solvent is preferably used at a temperature between room temperature and the boiling point of the solvent. Among these, it is preferable to extract using hot water.

  The rooibos extract can be purified and used as necessary, and the purification method is not particularly limited and can be appropriately selected. Examples include purification methods such as liquid-liquid partition extraction, various types of chromatography, membrane separation, and supercritical fluid extraction.

  For example, as the rooibos extract, a commercially available rooibos tea extract powder can be used. As a method for producing such a commercially available rooibos tea extract powder, the hot water extract of rooibos fermented young leaves is filtered, and the filtrate is used. A method of concentrating and drying to form a powder is known.

<Luteolin>
Luteolin is a component well known to be included in rooibos and is a compound having the following structure.

  Luteolin used in the present invention may be obtained from rooibos extract, may be obtained by chemical synthesis, or may be a commercially available product. The chemical synthesis method of luteolin is described in, for example, Liebigs Analen, Volume 1995, Issue 9, pages 1711-1515, 1995.

  The subject is ingested (administered) an effective amount of rooibos extract and / or luteolin. By “effective amount” is meant an amount that is significant for activating a muscarinic receptor or promoting salivary secretion in a subject. The significant amount can be appropriately set by those skilled in the art in consideration of the description of the examples in the present specification and the like. The significant amount usually depends on the condition and age of the subject.

  Since the agent or composition of the present invention can activate a muscarinic receptor, sweating promoting action, gastric acid secretion promoting action, urination promoting action, peristaltic movement promoting, saliva secretion promoting action and the like can be expected.

  In the case of dry powder of rooibos extract, the significant amount is, for example, preferably 1 mg / day to 10000 mg / day, more preferably 10 mg / day to 1000 mg / day, and still more preferably 20 mg / day to 500 mg / day. Such an amount may be taken (administered) once a day, or may be taken (administered) divided into a plurality of times per day. In the case of ingestion (administration) divided into multiple times, it is preferably 5 mg / dose to 50000 mg / dose, more preferably 50 mg / dose to 10000 mg / dose, more preferably 100 mg / dose to 5000 mg in terms of dry rooibos before extraction. / Times is even more preferred.

  In the case of luteolin, the significant amount is, for example, preferably 1 μg / day to 10,000 μg / day, more preferably 10 μg / day to 1000 μg / day, and even more preferably 20 μg / day to 500 μg / day. Such an amount may be taken (administered) once a day, or may be taken (administered) divided into a plurality of times per day. In the case of ingestion (administration) divided into a plurality of times, it is preferably 0.5 μg / time to 5000 μg / time, more preferably 5 μg / time to 500 μg / time, and even more preferably 10 μg / time to 250 μg / time.

  The ingestion (administration) route of the agent or composition of the present invention is not particularly limited as long as the desired effect of the present invention can be obtained. For example, oral (for example, intraoral, sublingual, etc.), parenteral (intravenous, intramuscular, subcutaneous, transdermal, nasal, transpulmonary, etc.) routes. Among these, a route with less invasiveness is preferable, and oral is more preferable.

  The intake (administration) of rooibos extract and / or luteolin is preferably by oral intake (oral administration).

  Examples of the rooibos extract and / or luteolin of the present invention in the case of oral ingestion (oral administration) include solid embodiments such as powder, fine granules, granules, capsules, sachets, tablets, boluses, lozenges, and films; Liquid forms such as aqueous solutions, extracts, suspensions, syrups, elixirs, emulsions, dispersions, and the like; semi-liquid forms, cream forms, paste forms, gel forms, and the like. Rooibos extract and / or luteolin can be ingested after being put in or dissolved in a liquid such as water or hot water (similar to drinking powdered tea) or in the form of a pill (powder or liquid in a capsule) It may be ingested (administered) in powder form or granule form (including freeze-dried granules).

  One aspect of the rooibos extract and / or luteolin of the present invention in the case of parenteral administration is, for example, an intravenous injection or an intramuscular injection such as an aqueous solution, extract, suspension, emulsion, dispersion or the like. Or subcutaneous injection; transdermal administration agent such as aqueous solution, extract, suspension, emulsion, dispersion, etc .; liquid, powder, fine particle, etc. of aqueous solution, extract, suspension, emulsion, dispersion, etc. Examples include nasal administration agents and transpulmonary administration agents.

  The agent of the present invention may be taken (administered) as it is. Moreover, you may use as an additive for providing at least 1 effect | action chosen from the group which consists of a muscarinic receptor activation effect | action and a saliva secretion promotion effect | action to food-drinks, a functional food, and a composition. Furthermore, the agent of the present invention can be included as an active ingredient in a composition for activating muscarinic receptors or promoting salivary secretion. As such a composition, for example, an oral composition, an internal use composition, and a pharmaceutical composition are preferable. When included as an active ingredient of the composition, at least one selected from the group consisting of rooibos extract and luteolin is contained in an amount of 0.00001 to 90% by mass (in the case of rooibos extract, converted to rooibos dry powder) The content is preferably 0.0001 to 50% by mass, more preferably 0.01 to 10% by mass.

  For foods and beverages, functional foods, and compositions that can be formulated with the agent of the present invention as an additive for imparting at least one action selected from the group consisting of a muscarinic receptor activating action and a salivary secretion promoting action, There are no particular restrictions, for example, beverages (soft drinks, carbonated drinks, nutritional drinks, powdered drinks, fruit drinks, milk drinks, jelly drinks, etc.), confectionery (cookies, cakes, gums, candy, tablets, gummies, buns, sheepskins) , Pudding, jelly, ice cream, sherbet, etc.), processed fishery products (kamaboko, chikuwa, hanpen, etc.), processed livestock products (hamburg, ham, sausage, winner, cheese, butter, yogurt, fresh cream, margarine, fermented milk, etc. ), Soup (powder soup, liquid soup, etc.), staple food (rice, noodles (dried noodles, raw noodles), bread, cereal, etc.) Seasonings (mayonnaise, shortening, dressing, sauce, sauce, soy sauce, etc.), toothpaste, liquid dentifrice, liquid dentifrice, dentifrice composition such as toothpaste, mouthwash composition, coating composition, Examples include oral pasta and mouth freshener composition.

  In addition to rooibos extract and / or luteolin, the agent or composition of the present invention may contain any conventional auxiliary ingredients used to produce solid or liquid dosage forms, such as excipients, dilutions. Agent, buffer agent, flavoring agent, coloring agent, flavoring agent, binder, surfactant, thickener, lubricant, suspending agent, preservative, antioxidant, abrasive, wetting agent, binder , One or more of pH adjusting agent, brightener, medicinal component, solvent, excipient and the like may be contained. Arbitrary auxiliary components can be appropriately selected depending on the dosage form. The components that can be blended in the present invention are not limited to these.

  The timing of ingesting (administering) the agent or composition of the present invention is not particularly limited, but since the agent or composition of the present invention has an immediate effect, it is ingested (administered) when a dry mouse is felt. It is preferable to do this.

  Hereinafter, the present invention will be described in detail based on examples, but the embodiments of the present invention are not limited to the examples.

[Example 1] Muscarinic receptor activation action Acetylcholine chloride (Wako Pure Chemical Industries, Ltd.) as a positive control, Rooibos extract (tea leaves 0.25% hot water extract, Marubeni Corporation), luteolin (as a ligand sample) EXTRASYNTHESE, Apiin (Applichem), Aspartin (Phytolab), Apigenin (EXTRASYNTHESE), Quercetin (EXTRASYNTHESE), Luteolin-7-O-Glucoside (Also: Synaroside, EXTRASYN ET-H-Eoline) , EXTRASYNTHESE), 4 ′, 6,7-trimethoxyisoflavone (EXTRASYNTHESE), 3 ′, 4 ′, 7-trimethoxyflavone (EXTRASYN) THESE), Daijin (Wako Pure Chemical Industries, Ltd.), Daizein (Wako Pure Chemical Industries, Ltd.), Genistein (Wako Pure Chemical Industries, Ltd.), Genistine (Wako Pure Chemical Industries, Ltd.), Glycine (Wako Pure Chemical Industries, Ltd.) Company), glycitein (Wako Pure Chemical Industries, Ltd.), ipriflavone (Wako Pure Chemical Industries, Ltd.), 5,7,4′-trimethoxyflavone (INDOFINE Chemical Company, Inc.).
Samples were suspended in 100% ethanol (Wako Pure Chemical Industries) except for the rooibos extract which had been subjected to hot water extraction, and diluted with a buffer attached to the assay kit. The cells were prepared by adding M3WT5 (ATCC) expressing mouse muscarinic M3 receptor to Ham's F-12 Nutrient Mix (Life Technologies) at a final concentration of 1% FBS at 2 × 10 ⁴ cells / 50 μl / It seed | inoculated to Biocoat Poly-D-Lysine 384-Well Black / Clear Plate (BD Biosciences) so that it might become a well. This was cultured overnight in a CO ₂ incubator and subjected to the assay.
In the assay, 42 μl of Fluo-4 Direct (registered trademark) Calcium Assay Kit (Life Technologies) was added per well, and the mixture was allowed to stand at room temperature for 30 minutes in the dark. Thereafter, using FDSS (Hamamatsu Photonics Co., Ltd.), 6 μl of atropine (Nippon Eye Drop Research Institute Co., Ltd.), which is an inhibitor of muscarinic M3 receptor, or HBSS-20 mM HEPES was added per well. Atropine was adjusted to a 10-fold concentration so that the final concentration was 10 μM. After adding an inhibitor or HEPES, the cells were cultured in a CO ₂ incubator for 10 minutes. After the culture, the rooibos extract was diluted with HBSS-20 mM HEPES to 5%, 0.5%, and 0.1%, respectively, and 12 μl of each was added using FDSS. For other samples, 12 μl of ligand or HBSS-20 mM HEPES 5 times higher than the final concentration was added per well, and measurement was performed for 3 minutes. Acetylcholine, which is a positive control, increased intracellular calcium from 10 nM, and suppression of the increase was confirmed by the coexistence of atropine. It was confirmed that the rooibos extract increased intracellular calcium ion in any sample, and the increase was greatly suppressed by adding atropine. Thus, when luteolin contained in the rooibos extract was subjected to the above assay system, it was confirmed that luteolin stimulated murine muscarinic M3 receptor as shown in FIG. This response was inhibited by the addition of atropine. On the other hand, in the flavonoid-related samples other than luteolin, the above response was not observed (FIGS. 2A to 2P).

[Example 2] Salivary secretion-promoting action in mouthwash 20 mL of an aqueous solution of 1% by weight rooibos extract powder (Rooibos tea extract powder MF, Maruzen Pharmaceutical Co., Ltd.) was included in the mouth and discharged after 30 seconds. Thereafter, the saliva was not swallowed at each time point, and the saliva accumulated in the oral cavity was discharged into the centrifuge tube. The amount of saliva secretion was measured by measuring the weight of the centrifuge tube. The control of this test is water, and n number is 12 persons.
The amount of saliva secretion during control (water) was set to 100%.
The results (average values) are shown in FIG.
From the results, it can be seen that the amount of saliva secretion increased when mouthwashed with the rooibos extract aqueous solution compared to when mouthwashed with the control.

[Example 3] Salivary secretion-promoting action in internal use Take 100 mg of rooibos extract powder (Rooibos tea dry extract F, Maruzen Pharmaceutical Co., Ltd.) with water and measure the amount of saliva secreted up to 2 hours after internal use. Measurement was carried out in the same manner as in the method.
Moreover, sensory evaluation by actual feeling evaluation was also performed according to the following evaluation criteria. The control of this test is water, and the n number is 5.
The results (average values) are shown in FIGS.
From the results, it can be seen that the amount of saliva secretion increased and the proportion of moisture in the mouth was higher when the rooibos extract was taken than when the control was taken.
<Evaluation criteria>
1 point: I felt a very moist feeling in my mouth.
2 points: I felt a feeling of moisture in my mouth.
3 points; I felt a little the feeling that moisture was given to the mouth.
4 points; The feeling of moisture in the mouth is neither.
5 points: I didn't feel much moisture in my mouth.
6 points; I didn't feel the moisture in my mouth.
7 points: I did not feel the feeling that moisture was given to my mouth.

Claims (6)

A muscarinic receptor M3 activator comprising at least one selected from the group consisting of an extract of Asparathus linearis and luteolin.   A salivary secretion promoter comprising at least one selected from the group consisting of an Asparathus linearis extract and luteolin. A composition for activating muscarinic receptor M3 or promoting saliva secretion, comprising as an active ingredient at least one selected from the group consisting of an extract of Asparathus linearis and luteolin.   The composition according to claim 3, comprising 1 to 50% by mass of at least one member selected from the group consisting of an extract of Asparathus linearis and luteolin. A food / beverage composition for activating muscarinic receptor M3 or for promoting salivation, comprising at least one selected from the group consisting of an extract of Asparathus linearis and luteolin. A food or drink for activating muscarinic receptor M3 or for promoting salivation, comprising at least one selected from the group consisting of an extract of Asparathus linearis and luteolin.

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