JP2021024858A - Hypotensive composition - Google Patents

Abstract

To provide a composition having the effect of improving blood pressure.SOLUTION: A hypotensive composition has extract from a ground part of a plant of Glycyrrhiza (Fabaceae).SELECTED DRAWING: None

Description

The present invention relates to a composition for lowering blood pressure containing an extract of a plant.

Hypertension is one of the typical lifestyle-related diseases, and the number of patients is increasing year by year. Hypertension is known to cause various disorders such as cerebral hemorrhage, cerebral infarction, heart failure, angina, myocardial infarction, and renal failure.

Substances that lower blood pressure have been searched for, and calcium antagonists, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor blockers, diuretics, sympathomimetics, and the like have been put into practical use. However, due to changes in consumer tastes in recent years, pharmaceuticals and foods with health claims containing naturally derived active ingredients have been demanded.

Licorice, which is made from a plant belonging to the genus Licorice of the family Leguminosae, has been used as a crude drug for a long time, and is known to have various actions such as antispasmodic, analgesic, antitussive, and expectorant.

The above-ground part of licorice (leaves and stems) was previously treated as waste because it cannot be used as a raw material for pharmaceuticals, but the extract from above-ground part of licorice may also have various effects. It has been known, and specifically, promotion of collagen production (Patent Document 1), improvement of bone metabolism (Patent Document 2), active oxygen scavenging and radical scavenging (Patent Document 3), hair cosmetics (Patent Document 4), For promotion of ceramide synthesis (Patent Document 5), suppression of increase in stem cell proliferation factor expression (Patent Document 6), suppression of increase in endoserine-1 mRNA expression (Patent Document 7), antiglycation (Patent Document 8), promotion of DKKI expression and promotion of DKKI expression Based on whitening (Patent Document 9), peroxysome proliferator-responsive receptor activation (Patent Document 10), and improvement of intestinal flora composition ratio, anti-chronic weak inflammation and maintenance of healthy obesity (Patent Document 11). Be done.

Further, a preparation for lowering blood pressure, etc. containing a licorice extract containing no glycyrrhizic acid (Patent Document 12), a composition for preventing or ameliorating hypertension, etc. containing a licorice hydrophobic extract (Patent Document 12). Patent Documents 13 and 14) are also known.

However, it has not been known so far that the extract from the above-ground part of licorice has an ACE inhibitory effect and a blood pressure lowering effect.

Japanese Unexamined Patent Publication No. 2000-191498 JP-A-2002-179585 JP-A-2003-081744 JP-A-2004-285018 Japanese Patent Application Laid-Open No. 2008-255051 Japanese Unexamined Patent Publication No. 2008-273875 JP-A-2010-215571 Japanese Unexamined Patent Publication No. 2012-121874 JP-A-2017-048155 JP-A-2009-242333 International Publication No. 2017/094892 JP-A-2002-114695 International Publication No. 2002/047699 International Publication No. 2008/143182

An object of the present invention is to provide a composition having a blood pressure improving effect such as lowering blood pressure. Another object of the present invention is to provide a food or pharmaceutical product containing the composition as an active ingredient for the prevention or alleviation of ACE or blood pressure-related diseases.

As a result of diligent studies, the present inventors have found that an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae has angiotensin converting enzyme (ACE) activity inhibitory action and blood pressure lowering action, and completes the present invention. It came to.
That is, the present invention includes the following aspects.
[1] A composition for lowering blood pressure, which comprises an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae.
[2] A composition having an angiotensin converting enzyme (ACE) inhibitory activity, which comprises an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae.
[3] The composition according to [1] or [2], wherein the above-ground part is a leaf and / or a stem.
[4] The composition according to any one of [1] to [3], wherein the extract of a plant belonging to the genus Licorice of the family Leguminosae is an extract extracted with water, an organic solvent, or a mixture thereof.
[5] The organic solvent is methanol, ethanol, 1-propanol, 2-propanol, acetone, 1-butanol, 2-butanol, 2-methyl-2-propanol, propylene glycol, glycerin, methyl acetate, ethyl acetate, hexane, At least one selected from the group consisting of diethyl ether, 2-butanol, cyclohexane, dichloromethane, edible fats and oils, propane, butane, 1,1,1,2-tetrafluoroethane and 1,1,2-trichloroethane. , [4].
[6] The composition according to any one of [1] to [5], wherein the concentration of the extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae is 0.20 mg / mL or more.
[7] The composition according to any one of [1] to [6], wherein the plant belonging to the genus Licorice of the family Leguminosae is a species of Ural licorice.
[8] The composition according to any one of [1] to [7], which is for eating and drinking.
[9] A food or pharmaceutical product containing the composition according to any one of [1] to [8] as an active ingredient for preventing or alleviating a disease related to blood pressure.
[10] A food or pharmaceutical product containing the composition according to any one of [1] to [8] as an active ingredient for preventing or alleviating angiotensin converting enzyme (ACE) -related diseases.

Since the composition of the present invention has an ACE inhibitory activity, a blood pressure lowering effect, and the like, it is possible to prevent or reduce ACE or a disease related to blood pressure. In addition, the composition of the present invention is highly safe because it contains a naturally derived extract as an active ingredient. In addition, the content of glycyrrhizic acid, which can cause an increase in blood pressure, is extremely low.
Therefore, the compositions of the present invention are useful as foods and pharmaceuticals for the prevention or alleviation of ACE or blood pressure related diseases.
In addition, in plants belonging to the genus Licorice of the family Leguminosae, parts (leaf and stem parts) that have not been previously available as raw materials for pharmaceuticals can also be effectively utilized.

The composition of the present invention contains an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae. Plants belonging to the genus Licorice of the family Licorice include, for example, Glycyrrhiza acanthocarpa, G.aspera, G.astragalina, G.bucharica, G.echinata (Russian licorice), G.eglandulosa, G.foetida, G.foetidissima, G.glabra. (Spanish licorice), G.gontscharovii, G.iconica, G.inflate (Shinki licorice), G.korshinskyi, G.lepidota (American licorice), G.pallidiflora, G.squamulosa, G.triphylla, G. uralensis (Licorice species), G.yunnanensis, etc. are mentioned, and varieties of plants belonging to the genus Licorice of the family Licorice can also be used. Licorice species and Glycyrrhiza uralensis species are preferable, and Glycyrrhiza uralensis species are more preferable. In the present invention, for example, Miyako licorice (also referred to as "Miyako No. 1") belonging to the genus Uralkanzo of the family Leguminosae is used.

The characteristics of Miyako licorice are "the plant height is quite high, the number of roots is quite large, the root thickness is quite thick, the root skin color is brown, the strength of the yellowish coloring of the cross section of the root is strong, and the stem Medium number, no strength of anthocyanin coloring on stem, no or coarse stem hair, long leaf length, fairly wide leaf width, medium wavy leaf strength, medium number of leaflets , Leaf blade length is quite long, leaflet blade width is quite wide, number of inflorescences is rather large, inflorescence length is medium, number of florets is quite large, flower color is N81A, dry The roots are fairly heavy and the glycyrrhizinic acid content is rather high "(color chart uses RHS). The distinction between "Miyako licorice" is recognized by the fact that the plant height is considerably higher and the leaf blade length of the leaflets is considerably longer than that of the "Northern agricultural trial system". In addition, the distinction is recognized by the fact that the plant height is considerably higher and the leaf blades of the leaflets are considerably longer than those of the "Hokkaido University". Furthermore, compared to the "medical system", the plant height is considerably higher, the leaf blades of the leaflets are considerably longer, and the early and late withering is late. Be done.

The extract contained in the composition of the present invention is, for example, an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae. The above-ground part of a plant belonging to the genus Licorice of the family Leguminosae includes, for example, parts such as leaves, stems, flowers, and seeds, preferably leaves and / or stems. That is, it may be an extract from either leaves or stems, or it may be an extract from leaves and stems. It is preferably an extract from the leaves and / or stems of plants belonging to the genus Licorice of the family Leguminosae, and more preferably an extract from the leaves and / or stems of licorice.

The extract in the present invention is obtained by extracting according to a conventional method using a solvent usually used for plant extraction. For example, it is obtained by immersing the plant in an extraction solvent and allowing it to stand or stir.

The plant used for extraction can be raw (raw), its dried product, or cut or crushed thereof.

Examples of the solvent used for extraction include solvents conventionally used for extracting plants, and examples thereof include water, organic solvents, and mixtures thereof.
Examples of the organic solvent include methanol, ethanol, 1-propanol, 2-propanol, acetone, 1-butanol, 2-butanol, 2-methyl-2-propanol, propylene glycol, glycerin, methyl acetate, ethyl acetate, hexane, and the like. Examples thereof include diethyl ether, 2-butanol, cyclohexane, dichloromethane, edible fats and oils, propane, butane, 1,1,1,2-tetrafluoroethane and 1,1,2-trichloroethane.
Preferred are water, ethanol or mixtures thereof. When a mixture of water and ethanol is used, the ethanol concentration in the mixture is, for example, 0.1 to 99.5 v / v%, preferably 5 to 80 v / v%, and more preferably 10 to 70 v / v%. Is.

The extraction method may be any commonly used method, and for example, extraction can be performed using any device at room temperature or under heating. Specifically, the plant can be put into a solvent-filled treatment layer, stirred occasionally to elute, and then centrifuged and filtered as necessary to remove the extraction residue to obtain an extract. it can. Further, as the extraction method, a method using a supercritical fluid composed of water, nitrous oxide, carbon dioxide or the like may be used.

The amount of the solvent used for extraction is not particularly limited, but is, for example, 1 to 100 mL, preferably 5 to 50 mL per 1 g of a plant (dry weight equivalent).
The extraction conditions are not particularly limited as long as they are sufficiently extracted, but usually, when the extraction temperature is low, extraction is performed for a longer period of time, and when the extraction solvent is high temperature, extraction is performed for a shorter time. The extraction temperature is appropriately set in the range of 0 to 100 ° C., preferably in the range of room temperature (about 25 ° C.) to the boiling point of the extraction solvent or less. The extraction time is set in the range of 10 minutes to 24 hours, preferably in the range of 30 minutes to 5 hours. When water is used as the extraction solvent, the extraction conditions are, for example, about 20 to 95 ° C. for about 30 minutes to 3 hours. When ethanol or a mixed solvent of water and ethanol is used as the extraction solvent, the extraction conditions are, for example, about 20 to 80 ° C. for about 30 minutes to 3 hours.
The number of extractions may be one, or may be multiple to increase the yield.

The obtained extract can be used as it is as the composition of the present invention. In addition, the extract may be subjected to purification for the purpose of decolorization or deodorization, if necessary. Examples of the purification method include activated carbon treatment, adsorption treatment, ion exchange resin treatment and the like.

The extract may be further concentrated or concentrated to dryness, diluted with a suitable solvent, or optionally added excipients and then dried to a powder. Examples of the method for concentration or concentration to dryness include vacuum concentration (method using an evaporator, etc.) and membrane concentration. Examples of the powdering method include spray drying, freeze drying, and fluidized bed drying. Examples of the excipient used when the extract is powdered include those conventionally used, and examples thereof include modified starch and starch decomposition products (for example, dextrin and maltodextrin).

The "extract" in the present invention includes various solvent extracts obtained by the above-mentioned extraction method, its purified fraction, its diluted solution, its concentrated solution, and its dried product (powder, etc.).

The amount of the extract from the above-ground part of the plant belonging to the genus Licorice of the family Leguminosae contained in the composition of the present invention is not particularly limited as long as it can exert a desired effect. For example, when the composition of the present invention is a liquid composition, the lower limit of the concentration in the liquid composition (dry weight of extract / volume of liquid composition) is, for example, 0.01 mg / mL or more, preferably 0.01 mg / mL or more. It is 0.10 mg / mL or more, and the upper limit thereof is, for example, 100 mg / mL or less, preferably 10 mg / mL or less.
When the composition of the present invention is a solid composition, the lower limit of the blending ratio (dry weight of extract / dry weight of composition) in the solid composition is preferably 0.01% by weight or more, for example. Is 0.1% by weight or more, and the upper limit thereof is, for example, 99% by weight or less, preferably 50% by weight or less.

The extract in the present invention has an angiotensin converting enzyme (ACE) activity inhibitory action. ACE produces angiotensin II having a vasoconstrictive effect in the renin-angiotensin system, which is a pressor system, and is also a degrading enzyme of bradykinin having a hypotensive effect. Therefore, inhibiting the ACE activity produces an antihypertensive effect by two mechanisms: suppression of angiotensin II production and suppression of bradykinin inactivation.
Therefore, the composition of the present invention containing the extract exhibits an ACE activity inhibitory action and is useful for improving blood pressure, particularly lowering blood pressure.

"Blood pressure improvement" here refers to improving vasodilation in mammals such as humans, maintaining normal vasodilation, lowering blood pressure, maintaining stable blood pressure, and protecting vascular endothelial cells from degeneration. Includes things to do. Vasodilation also means dilation of blood vessels, especially dilation of arterioles that results in increased blood flow to parts of the blood vessels.

Moreover, since the extract in the present invention is derived from a plant belonging to the genus Licorice of the family Leguminosae, it is highly safe. Therefore, the compositions of the present invention are useful as foods and pharmaceuticals, especially as foods and pharmaceuticals for the prevention or alleviation of diseases associated with blood pressure or ACE.

The pharmaceutical product of the present invention is produced according to a conventional method using the above extract or the above composition. For example, tablets, capsules, granules, etc., using known methods such as adding and mixing the extract or composition with a pharmaceutically acceptable carrier or additive commonly used in the field of formulation technology. It can be in the form of a powder, oral solution, syrup, etc. for oral administration.
Examples of the carrier or additive include excipients, disintegrants, binders, fluidizers, lubricants, colorants, pH regulators, surfactants, stabilizers, acidulants, fragrances and the like. These additives are used in conventional amounts in the field of formulation technology.

Excipients include, for example, starches such as corn starch, potato starch, wheat costarch, rice starch, partially pregelatinized starch, pregelatinized starch, perforated starch; lactose hydrate, sucrose, fructose, glucose, mannitol. , Sorbitol, Erythritol, Xylitol, Trehalose, Martitol, Powdered Reduced Starch Sugar Water Ame, sugar alcohols such as lactitol; anhydrous calcium hydrogen phosphate, crystalline cellulose, powdered cellulose, precipitated calcium carbonate, calcium carbonate and the like.
Examples of the disintegrant include carmellose, carmellose calcium, carboxymethyl starch sodium, carboxymethyl cellulose, carboxymethyl cellulose calcium, carboxymethyl starch sodium, croscarmellose sodium, crospovidone, low degree of substitution hydroxypropyl cellulose (L-HPC), and the like. Hydroxypropyl starch or the like is used, preferably croscarmellose sodium, L-HPC.
Examples of the binder include hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinylpyrrolidone, copolyvidone, gum arabic powder, methyl cellulose, low-substituted hydroxypropyl cellulose, hypromellose, sodium carmellose, dextrin, partially pregelatinized starch, purulan, and gum arabic. , Canten, gelatin, tragant, sodium alginate and the like are used, preferably hydroxypropyl cellulose, hydroxypropyl methyl cellulose and the like.
As the fluidizing agent, for example, light anhydrous silicic acid, hydrous silicon dioxide, calcium silicate, magnesium silicate, magnesium aluminometasilicate, talc and the like are used, and preferably light anhydrous silicic acid and magnesium aluminometasilicate. Is.
Examples of the lubricant include stearic acid, magnesium stearate, calcium stearate, talc, and sucrose fatty acid ester.
Coloring agents include, for example, yellow iron sesquioxide, iron sesquioxide, edible blue No. 1, edible blue No. 2, edible yellow No. 4, edible yellow No. 5, edible green No. 3, edible red No. 2, and edible red No. 3. , Edible red No. 102, edible red No. 104, edible red No. 105, edible red No. 106, edible lake pigment, riboflavin, sodium riboflavin phosphate and the like.
Examples of the pH adjuster include citric acid, phosphoric acid, carbonic acid, tartaric acid, fumaric acid, acetic acid, amino acids and salts thereof.
Examples of the surfactant include sodium lauryl sulfate, polysorbate 80, polyoxyethylene (160) polyoxypropylene (30) glycol and the like.
Examples of the stabilizer include tocopherol, tetrasodium edetate, nicotinamide, cyclodextrins and the like.
Examples of the acidulant include ascorbic acid, citric acid, tartaric acid, malic acid and the like.
Examples of the fragrance include L-menthol, peppermint oil, lemon oil, vanillin and the like.
As the above-mentioned carrier or additive, two or more kinds may be appropriately mixed and used.

The pharmaceutical product of the present invention is preferably in the form of a unit dose containing 0.05 to 5 g, preferably 0.1 to 1.5 g of the extract in terms of dry weight. Here, the "unit dose form" means physically divided units, and each unit contains a predetermined amount of active ingredient calculated so as to obtain a desired effect together with a carrier or an additive. There is.

The dose of the drug of the present invention varies depending on the age, sex, body weight and condition of the subject to be administered, the route of administration, the administration schedule, the dosage form and the composition of the preparation, and is, for example, 1 to 1000 mg, preferably 5 to 500 mg. The dose may be administered once a day, or in two or three or more divided doses.

The food product of the present invention is produced according to a conventional method using the above extract or the above composition. For example, it is produced by a known means such as adding the extract or the composition to a food product, a food material thereof, or an intermediate material for the food product. For example, it is produced by extracting the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae with water or ethanol, concentrating it, adding the obtained concentrated extract to a food material, and then making it into a food by a conventional method.

Examples of the above-mentioned foods include confectionery (for example, snack confectionery such as potato chips, baked confectionery such as biscuits or cookies, chocolate, gum, candy, etc.), desserts (for example, pudding, jelly, yogurt, ice cream, etc.). In addition to favorite foods such as, noodles (eg buckwheat, udon, ramen or pasta, etc.), foods similar to main foods such as serial foods (eg corn flakes or oatmeal, etc.), seasoned foods (eg soup, curry or stew, etc.) , Processed agricultural products (eg jam, etc.), milk oil foods (eg spreads or cheese, etc.), health foods (eg protein or fiber, etc.), calorie-adjusted foods, non-alcoholic beverages (eg, roasted soybean tea beverages, grain tea, coffee beverages) , Tea drinks, green tea drinks, barley tea drinks, matcha drinks, vegetable juice drinks, orange juice, grapefruit juice, soft drinks, etc., or alcoholic drinks (eg beer, wine, sake, sparkling liquor, plum wine, whiskey, brandy, shochu, etc. Vocabulary, lamb, gin, liqueurs or cocktails, etc.).

The food of the present invention can be, for example, general foods including nutritional supplements, health supplements, nutritionally adjusted foods, etc., and health functional foods including foods for specified health use, nutritional functional foods and foods with functional claims. For example, the food of the present invention is based on scientific evidence that a specific health purpose can be expected (for example, "the function of lowering blood pressure has been reported and it is suitable for people with high blood pressure"). It can be a food with functional claims to be labeled. In addition, the food product of the present invention may be a supplement having a form for oral ingestion such as tablets, capsules, powders, granules, liquids and syrups.

The amount of the extract contained in the food of the present invention may be an amount capable of exerting a desired effect, and may be appropriately determined according to the general intake of the food, the type of the food, the composition and the like. For example, the food product of the present invention can contain the extract in the range of 0.1 to 99.0% by weight as a dry weight.

The amount of the food of the present invention may vary depending on the age, sex, body weight and condition of the subject to be ingested, the type and composition of the food, and the like, and is, for example, 1 to 1000 mg, preferably 5 to 500 mg. The intake may be taken once a day, or divided into two or three or more times a day.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

[Examples 1 to 5] Preparation of water-based extract from leaves and stems of licorice, about 300 L of water is added to 18 kg of leaves and stems of licorice, extracted at 90 ° C. for about 2 hours, mesh-filtered, and 250 L. An extract was obtained. Further, this extract was concentrated under reduced pressure at about 60 ° C. to obtain 9.7 kg of a concentrated solution. 280 g of a starch decomposition product (trade name: Max 1000, manufacturer: Matsutani Chemical Industry Co., Ltd.) was dissolved in this concentrate as an excipient, and then 2092 g was spray-dried to obtain 498 g of a dry extract.
Approximately 1.0 g of the above dry extract is extracted with 20 mL of a 50% ethanol solution, diluted appropriately with 0.1 mol / L HEPES buffer (pH 8.3), and 0.26 mg / mL (Example 1), 0. .28 mg / mL (Example 2), 0.30 mg / mL (Example 3), 0.34 mg / mL (Example 4) and 0.40 mg / mL (Example 5) of Tokyo licorice with sample concentrations. Water extract-containing compositions from leaves and stems were obtained.

[Examples 6 to 10] Preparation of ethanol extract from leaves and stems of Tokyo licorice Add about 300 L of 30% ethanol to 18 kg of leaves and stems of Tokyo licorice, extract at 60 ° C. for about 2 hours, filter with mesh, and 250 L. Extract was obtained. Further, this extract was concentrated under reduced pressure at about 60 ° C. to obtain 10 kg of a concentrated solution. 200 g of a starch decomposition product (trade name: Max 1000, manufacturer: Matsutani Chemical Industry Co., Ltd.) was dissolved in this concentrate as an excipient, and then 2928 g was spray-dried to obtain 320 g of a dry extract.
Approximately 1.0 g of the above dry extract is extracted with 20 mL of a 50% ethanol solution and then appropriately diluted with 0.1 mol / L HEPES buffer (pH 8.3) to 0.25 mg / mL (Example 6), 0. .35 mg / mL (Example 7), 0.45 mg / mL (Example 8), 0.55 mg / mL (Example 9) and 0.65 mg / mL (Example 10) of Tokyo licorice with sample concentrations. An ethanol extract-containing composition from leaves and stems was obtained.

[Reference Examples 1 to 5]
Sardenpeptide (trade name: Sardenpeptide N100, manufacturer: Senmi Extract Co., Ltd.) is appropriately diluted with 0.1 mol / L HEPES buffer (pH 8.3) and 0.1 mg / mL (reference). Sample concentrations of Example 1), 0.2 mg / mL (Reference Example 2), 0.5 mg / mL (Reference Example 3), 0.7 mg / mL (Reference Example 4) and 0.9 mg / mL (Reference Example 5) A sardine peptide-containing composition having the above was obtained.

[Test Example 1]
1. 1. Test outline Angiotensin converting enzyme (ACE) activity test is performed by Nakano et al. Biosci Biotechnol. Biochem. , 70, 1118-1126 (2006), after fluorescentizing a dipeptide that is degraded by ACE from a substrate (His-His-Leu) with orthophthalaldehyde (hereinafter abbreviated as "OPA"). This was carried out by measuring the fluorescence intensity of the reaction product. Inhibition of ACE activity was evaluated based on the relative ACE activity when the activity of the untreated group to which the test solution was not added was set to 100%. IC ₅₀ values were calculated from the graph of sample concentration (mg / mL) and inhibition rates (%).

2. 2. Specimens The compositions obtained in Examples 1 to 10 above were used as the test solutions of the following samples 1) and 2). Further, as a positive control (sample 3)), the compositions obtained in Reference Examples 1 to 5 above were used.
1) Water extract from leaves and stems of Tokyo licorice (Examples 1 to 5)
2) Ethanol extract from leaves and stems of Tokyo licorice (Examples 6 to 10)
3) Sarden peptide (Reference Examples 1 to 5)

3. 3. Test Method 25 μL of 0.1 mol / L HEPES buffer (pH 8.3) (untreated group) or each test solution was added to a 96-well microplate, 25 μL of 13 mU / mL ACE solution was added, and the mixture was incubated at 37 ° C. for 5 minutes. 25 μL of an 8 mmol / L His-His-Leu solution was added and reacted at 37 ° C. for 30 minutes. Then, 25 μL of 0.1 mol / L sodium hydroxide solution was added to stop the reaction, 25 μL of 1% OPA solution was added, and the mixture was left at room temperature for 20 minutes. Further, 25 μL of 0.1 mol / L hydrochloric acid was added and left at room temperature for 10 minutes, and the fluorescence intensity was measured with a microplate reader. The blank was similarly tested using PBS instead of 13 mU / mL ACE solution.
<Microplate reader operating conditions>
Model: SH-9000 Lab
Measurement conditions: Fluorescence, endpoint mode, bottom lead Excitation wavelength: 355 nm
Fluorescence wavelength: 460 nm

4. Test Results Tables 1 to 3 show the results of the inhibition rate in each test solution. As a result, concentration-dependent enhancement of ACE activity inhibition was observed. The IC ₅₀ values calculated from the graph showing the relationship between sample concentration and inhibition rates shown in Table 4.

[Examples 11 and 12] Preparation of water-based extract from above-ground part of Tokyo licorice Add about 300 L of water to 18 kg of leaves and stems of Tokyo licorice, extract at 90 ° C. for about 2 hours, then mesh filter to extract 250 L. I got the liquid. Further, this extract was concentrated under reduced pressure at about 60 ° C. to obtain 9.7 kg of a concentrated solution. 280 g of a starch decomposition product (trade name: Max 1000, manufacturer: Matsutani Chemical Industry Co., Ltd.) was dissolved in this concentrate as an excipient, then 2092 g was spray-dried, and 498 g of a dry extract (Examples 11 and 12). Got

[Reference Example 6] Preparation of water-based extract from the underground part of Tokyo licorice Add about 30 L of water to 3 kg of the root and stron of Tokyo licorice, extract at 90 ° C for about 2 hours, filter with a mesh, and extract 23.2 L. Got Further, this extract was concentrated under reduced pressure at about 60 ° C. to obtain 2544 g of a concentrated solution. 2511 g of this concentrate was spray-dried to obtain 608 g of a dry extract (Reference Example 6).

[Test Example 2]
Example 11 (100 mg / kg), Example 12 (1000 mg / kg), Reference Example 6 (1000 mg / kg), Reference Example 7 (positive control, Sardenpeptide (commodity)) in 14-week-old SHR / NCrlCrlj male rats. Name: Sardenpeptide N100, Manufacturer: Senmi Extract Co., Ltd., 1000 mg / kg), Reference Example 8 (Positive control, Captopril (Manufacturer: Tokyo Kasei Kogyo Co., Ltd.), 60 mg / kg) was administered as a single dose. The systolic blood pressure (SBP) was evaluated. The control group received the vehicle (water for injection) at a dose of 10 mL / kg. SBP was performed using a non-invasive blood pressure monitor (BP MONITOR FOR MICE & RATS, Muromachi Machinery, MODEL MK-2000) before administration (before grouping) of each administered substance and 2, 4, 6 and 8 hours after administration. It was measured.

The results are shown in Table 5. In the water extract (Examples 11 and 12) administration group of the above-ground part of Tokyo licorice, a significant decrease was observed in SBP 2 and 8 hours after administration of 100 mg / kg and 2 hours after administration of 1000 mg / kg as compared with the control group. Was done. In addition, in the water extract (Reference Example 6) administration group in the underground part of Tokyo licorice, a significant decrease was observed in SBP 2 hours after administration as compared with the control group. In the Sarden peptide-administered group, a significant decrease in SBP 6 and 8 hours after administration was observed as compared with the control group. From the above results, it was suggested that the above-ground licorice extract, the underground licorice extract, and the sardine peptide have an inhibitory effect on the increase in blood pressure in spontaneously hypertensive rats.
N: Number of rats, each value is expressed as mean ± SE.
#: P <0.05, ##: P <0.01: Significant difference from the control group (Danette's multiple comparison test)
*: P <0.05, **: P <0.01: Significant difference from the control group (Student's t-test)

Since the composition of the present invention has an ACE inhibitory activity, a blood pressure lowering effect, and the like, it is possible to prevent or reduce ACE or a disease related to blood pressure. In addition, the composition of the present invention is highly safe because it contains a naturally derived extract as an active ingredient. Furthermore, it has been confirmed that the content of glycyrrhizic acid, which causes pseudoaldosteronism that causes an increase in blood pressure, is extremely low.
Therefore, the compositions of the present invention can be used as foods and pharmaceuticals for the prevention or alleviation of diseases related to ACE or blood pressure.
In addition, in plants belonging to the genus Licorice of the family Leguminosae, parts (leaf and stem parts) that have not been previously available as raw materials for pharmaceuticals can also be effectively utilized.

Claims (10)

A composition for lowering blood pressure, which comprises an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae. A composition having an angiotensin converting enzyme (ACE) inhibitory activity, which comprises an extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae. The composition according to claim 1 or 2, wherein the above-ground part is a leaf and / or a stem. The composition according to any one of claims 1 to 3, wherein the extract of a plant belonging to the genus Licorice of the family Leguminosae is an extract extracted with water, an organic solvent, or a mixture thereof. Organic solvents are methanol, ethanol, 1-propanol, 2-propanol, acetone, 1-butanol, 2-butanol, 2-methyl-2-propanol, propylene glycol, glycerin, methyl acetate, ethyl acetate, hexane, diethyl ether, Claim that it is at least one selected from the group consisting of 2-butanol, cyclohexane, dichloromethane, edible fats and oils, propane, butane, 1,1,1,2-tetrafluoroethane and 1,1,2-trichloroethane. The composition according to 4. The composition according to any one of claims 1 to 5, wherein the concentration of the extract from the above-ground part of a plant belonging to the genus Licorice of the family Leguminosae is 0.20 mg / mL or more. The composition according to any one of claims 1 to 6, wherein the plant belonging to the genus Licorice of the family Leguminosae is a species of Ural licorice. The composition according to any one of claims 1 to 7, which is for eating and drinking. A food or pharmaceutical product containing the composition according to any one of claims 1 to 8 as an active ingredient for preventing or alleviating a disease related to blood pressure. A food or pharmaceutical product containing the composition according to any one of claims 1 to 8 as an active ingredient for preventing or alleviating angiotensin converting enzyme (ACE) -related diseases.