KR101847452B1 - Anti-hypertensive composition comprising Scapharca broughtonii extract as effective component - Google Patents

Abstract

The present invention relates to an anti-hypertension composition containing a Scapharca broughtonii extract as an active ingredient. The Scapharca broughtonii extract of the present invention, more specifically, the Scapharca broughtonii solvent extract or the Scapharca broughtonii hydrolyzed extract has an excellent effect of inhibiting activity of an angiotensin I converting enzyme. In particular, the Scapharca broughtonii hydrolyzed extract is more effective in inhibiting the activity of the angiotensin I converting enzyme than the Scapharca broughtonii solvent extract. Therefore, the Scapharca broughtonii extract of the present invention, more specifically, the Scapharca broughtonii hydrolyzed extract can be used for an anti-hypertension health functional food or a therapeutic agent for hypertension.

Description

[0001] The present invention relates to an antihypertensive composition comprising Scapharca broughtonii extract as an active ingredient,

The present invention relates to a composition for anti-hypertension containing an arcanoid extract as an active ingredient.

Hypertension is a chronic degenerative disease of the circulatory system and is explained by the physiological mechanism of the renin-angiotensin system, which plays an important role in regulating blood pressure and electrolyte balance. Angiotensin-I converting enzyme (ACE) converts angiotensin II, which has vasoconstrictive action, by digesting dipeptide (His-Leu) from angiotensin I, a decapeptide. The increase of angiotensin II produced by angiotensin converting enzyme promotes hypertension by promoting strong hypertension and secretion of aldosterone, an antidiuretic hormone, and inhibiting the excretion of water and sodium to increase circulating blood volume. In addition, angiotensin-converting enzyme degrades and inactivates blood-relaxing bradykinin, resulting in a rise in blood pressure. Therefore, inhibition of the activity of angiotensin converting enzyme inhibits vasoconstriction and blood pressure lowering effect can be shown to lower the hypertension.

Antihypertensive agents for hypertension include diuretics, beta-blockers, sympathomimetics, vasodilators, Ca-antagonists and AT1-receptor blockers. However, most of these drugs cause side effects such as elevated blood viscosity, electrolyte imbalance, diabetes induction, worse heart failure, dysfunction, and kidney dysfunction.

On the other hand, as interest in health and longevity has been concentrated in recent years, various physiological activities such as anticancer, anti-aging and anti-hypertension, which are represented by food ingredients, are being carried out variously. Substances derived from food are of great value in terms of their natural nature.

On the other hand, ark shell (Scapharca broughtonii ) is a shellfish belonging to the eustachian clam, and its shape is ovate, and both shells are swollen well and thinnest among the eustachian membranes. On the outer surface of the shell, there are 42 ~ 43 pairs of white, convex rhyolite, and there is a dark brown skin on the shell. The inside of the shell is white, and it is a large shell with a shell length of 120 mm, an angle of 90 mm, and a width of 75 mm. It is widely distributed in the southern coast of Korea and the eastern coast, and lives mainly from the vicinity of the Korean peninsula to a depth of 50m.

Archeopteryus is a life style, eggs arising in summer are modified in seawater, and after 2 ~ 3 weeks of floating life, they go into attachment life by using swam. The attachment life is about 2 months and depends on the environment of the place where it is attached. After attaching life period, it infiltrates in low quality and lives.

The arboreal larvae are mainly eaten raw, but recently they started to be used as raw materials for canning. The total calories per 100g of the raw shellfish are 81 ㎈, and the composition is 79.8g of moisture, 15.5g of protein, 0.5g of fat, 3.5g of carbohydrate and 0.7g of ash, and vitamin A, B1, B2C and niacin ), And it is known to be effective for preventing anemia, improving digestive function, protecting eyesight, relieving fatigue, liver detoxification, and maintaining bone density. However, there is little research on antihypertensive efficacy.

Meanwhile. Korean Patent No. 1533308 discloses a pharmaceutical composition for the prevention or treatment of cardiovascular diseases, which comprises, as an active ingredient, a peptide exhibiting an angiotensin-I converting enzyme inhibitory activity, which is isolated from fractions of oyster enzyme hydrolyzate, There has not been disclosed a composition for anti-hypertension containing an extract as an active ingredient.

The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide an antihypertensive composition containing an arthropod extract as an active ingredient, and confirming that the composition inhibits the activity of angiotensin I converting enzyme, thereby completing the present invention .

In order to solve the above problems, the present invention provides a health functional food composition for preventing or ameliorating hypertension containing an arcanoid extract as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating hypertension comprising an arcanoid extract as an active ingredient.

In particular, the shell extract or the shell hydrolyzed extract of the present invention has an excellent effect of inhibiting the activity of angiotensin I converting enzyme, and in particular, the shell hydrolyzate extract has an activity of angiotensin I converting enzyme The arthropod extract of the present invention, in particular, the hydrolyzed extract of shellfish, can be used as a material for a health functional food composition for antihypertensive or a pharmaceutical composition for the prevention or treatment of hypertension.

Fig. 1 shows the results of confirming the inhibitory effect of arcuate solvent extract on angiotensin I converting enzyme activity. Captopril is a positive control.
FIG. 2 shows the results of confirming the inhibitory effect of the hydrolyzate of the shellfish on angiotensin I converting enzyme activity. Captopril is a positive control.

The present invention relates to a health functional food composition for preventing or ameliorating hypertension containing an arthritic ginseng extract as an active ingredient.

The above-mentioned shellfish extract is extracted by solvent extraction or hydrolysis, but is not limited thereto.

In one embodiment of the present invention, the solvent extract is water, C ₁ -C ₄ lower alcohol, or a mixture thereof, as a solvent. Preferably, water is used as a solvent, but the present invention is not limited thereto.

The hydrolysis is hydrolysis using a protein hydrolyzing enzyme, and the protein hydrolyzing enzyme is Protamex and Neutrase, but is not limited thereto.

The addition of the hydrolytic enzyme is preferably sequential addition, but not limited thereto, of inactivating the protease after the reaction, and then adding Ntrease.

The protamex of the present invention is a protease of bacillus sp. , Nutraceutical is Bacillus amyloliquefaciens ( Bacillus sp. Amyloliquefaciens ) is a protease of bacteria.

In one embodiment of the present invention,

1) adding water and protamase to the demineralized crushed product of the deionized water to perform primary hydrolysis, and then deactivating the protamex;

2) dehydrogenating the protease by inactivating the protease by inactivating the protease by adding the protease to the hydrolyzate, and then deactivating the protease;

3) filtering the hydrolyzate inactivated by the Ntrease of step 2), adding ethanol and centrifuging to obtain supernatant; And

4) Concentrating the supernatant of step 4) under reduced pressure and freeze-drying,

More preferably,

1) To 100 g of demineralized crushed material, 1 l of water and protamase were added and the mixture was subjected to primary hydrolysis at 35 to 45 ° C for 2 to 4 hours, followed by hydrolysis at 90 to 100 ° C for 20 to 40 minutes. Deactivating the mage;

2) Nutraceuticals are added to the hydrolyzate obtained by inactivating the protamase of step 1), and then subjected to secondary hydrolysis at 45 to 55 ° C for 2 to 4 hours, followed by hydrolysis at 90 to 100 ° C for 20 to 40 minutes Deactivating the Ntra agent;

3) filtration of the hydrolyzate inactivated by the Ntrease in step 2), followed by adding 60% (v / v) ethanol of 2 parts skin to 1 part skin of the filtrate and centrifuging to obtain supernatant; And

4) concentrating the supernatant of step 4) under reduced pressure, followed by freeze-drying, but the present invention is not limited thereto.

In one embodiment of the present invention, the arboresa extract inhibits the activity of the angiotensin converting enzyme (ACE).

The health functional food composition of the present invention can be prepared into any one form selected from powder, granule, ring, tablet, capsule, candy, syrup foam, and beverage. However, it can be formulated to prevent or improve hypertension Is not particularly limited.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method.

The active ingredient may be suitably used depending on its intended use (prevention or improvement). Generally, in the production of food or beverage, it is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the health functional food composition of the present invention. However, in the case of long-term intake for the purpose of health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of foods to which the health functional food composition can be added include meat products, sausages, breads, chocolate, candies, snacks, confectionery, pizza, ramen noodles, other noodles, gums, dairy products including ice cream, various soups, Drinks, and vitamin complexes, all of which include healthy foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when it is made of a drink, it may contain, in addition to the active ingredient, a natural carbohydrate or a flavoring agent as an additional ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above-mentioned health functional food composition, it is possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like.

The present invention also relates to a pharmaceutical composition for preventing or treating hypertension containing an arthritic ginseng extract as an active ingredient.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of pharmaceutical compositions. The pharmaceutical dosage forms of the compositions according to the invention may be used alone or in combination with other pharmaceutically active compounds as well as in suitable combinations.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and injections according to conventional methods . Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include withexol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Examples. It is to be understood by those skilled in the art that these preparations and examples are merely intended to explain the present invention more specifically and that the scope of the present invention is not limited thereto.

Manufacturing example  1. Preparation of Ark shell extract

Archeopteryx ( Scapharca broughtonii ) was purchased from Byeokgyo-eup (Bugyo-eup), Bogyeong -eup (Bugyo-eup), and collected in Bucheon-eup, Boseong-gun, Jeollanam-do in December 2015. After frozen in 1kg unit, it was kept at -40 ℃ until it was used for experiment.

The raw materials of the stored shellfish were thawed for about 4 hours at room temperature before extraction and desalted by rapid washing three times to tap water of about 10 times (w / v) compared to the shellfish. The sodium chloride content was measured by self-inspection Respectively. The desalted raw material was used as an off-angle.

1) Manufacture of shellfish hot water extract

After desalting, the arsenic was crushed, and 100 g of the crushed material was immersed in 1 L of distilled water, and the mixture was placed in a 2 L reflux condenser and extracted with a heating mantle (Tops, MS-E106) for 3 hours at 100 ° C . After completion of the extraction, the mixture was filtered using a filter paper (Watman No. 2), and the filtrate was lyophilized in a freeze dryer (FDS8508. Ilshin biobase, Korea) and used as an experimental sample.

2) Preparation of ethanol extract of shellfish

100 g of the pulverized product was immersed in 1 L of 30% (v / v) ethanol or 100% (v / v) ethanol, and the mixture was shaken in a shaking incubator (DA- SIM-T, Dong a, Korea) at 120 rpm for 3 hours. After completion of the extraction, the mixture was filtered using a filter paper (Watman No. 2), and then the organic solvent was removed on a rotary evaporator (N-100v-W, Eyela, japan) and the water layer was transferred to a lyophilizer (FDS8508. ) And used as a sample of 30% (v / v) ethanol extract or 100% (v / v) ethanol extract, respectively.

3) Preparation of the hydrolyzed extract of the arborescens

After desalting, the arched shell was crushed, and 100 g of the pulverized material was placed in a 2 L round-bottomed media vial and immersed in 1 L of distilled water. Protamex (1.5AU / g) was added to the immersed sample to 1% (w / v) and reacted in a constant temperature bath (BF-10BF, biofree) at 40 ℃ for 3 hours. The primary enzyme hydrolyzate of the procambium treated with protamase was reacted at 100 ° C for 30 minutes using an autoclave (AC-14, JEIO TECH, Korea) to inactivate the enzyme. Then, 0.81 ml of neutrase was added to 1 ml of the enzyme-inactivated primary enzyme hydrolyzate, and the mixture was reacted in a constant temperature bath (BF-10BF, biofree) at 50 ° C for 3 hours. , JEIO TECH, Korea) for 30 minutes at 100 ° C to inactivate the enzyme. After filtration using a filter paper (Watman No. 2), 60% (v / v) ethanol of 2 parts skin was added to 1 part skin of the filtrate and centrifuged (3000 × g, 10 minutes). After centrifugation, the supernatant was collected and concentrated under reduced pressure. The organic solvent was removed, and the extract was lyophilized using a freeze dryer (FDS8508. Ilshin biobase, Korea) and used as a sample.

Example  One. Angiotensin  Ⅰ Converting enzyme inhibitory activity

Angiotensin I converting enzyme inhibitory activity was measured using an ACE kit-WST (A502, Dojindo, Japan). The enzyme and substrate buffer provided in the kit were used, and the inhibitory activity was measured according to the experimental method of the kit.

To the enzyme A and the enzyme B in the kit, 2 ml of each of the third distilled water was added, and 1.8 ml of the prepared enzyme solution was added to the enzyme solution. The enzyme working solution was prepared.

The indicator working solution was prepared by adding 3 ml of the third distilled water to the enzyme C and the crude enzyme solution, 2.8 ml of the prepared enzyme solution, and mixing the solution with 5 ml of the indicator solution before measuring the absorbance .

The angiotensin I converting enzyme inhibitory activity was determined by adding 20 μl of a substrate buffer and 20 μl of an enzyme working solution to 20 μl of each sample of Preparation Example 1 and incubating the cells at 37 ° C. for 1 hour in an incubator Then, 200 μl of indicator working solution was added and reacted at room temperature for 10 minutes. Absorbance was measured at 450 nm using a microplate ELISA reader, and the value was substituted into the following formula 1 . Captopril (captopril, Sigma, USA) was used as a positive control. The preparation methods of the sample addition group, blank 1 and blank 2 are shown in Table 1 below.

[Equation 1]

(Inhibition rate%) = [(absorbance of blank 1 - absorbance of group with added sample) / (absorbance of blank 1 - absorbance of blank 2)] × 100

Preparation method (l) of sample addition group, blank1 and blank 2, Sample addition group blank1 blank 2 The sample of Preparation Example 1 20 - - Distilled water - 20 40 Substrate buffer 20 20 20 Enzyme working solution 20 20 - Indicator working solution 200 200 200 total 260 260 260

As a result, as shown in FIG. 1 and FIG. 2, the arcanum extract inhibited the activity of angiotensin I converting enzyme, and the inhibition efficiency was different depending on the solvent or preparation method of the extract.

The inhibitory effect of hydrothermal extract on angiotensin Ⅰ conversion enzyme activity was better than ethanol extract of shellfish, and the inhibition rate of angiotensin I converting enzyme activity was more remarkable than that of water or ethanol extracts when the shell hydrolysis extract was treated. The hydrolyzate of the shellfish showed inhibition of angiotensin I converting enzyme activity in a dose - dependent manner.

It was confirmed that the arthropod extract, especially the shell hydrolyzed extract, had an antihypertensive effect by inhibiting the activity of angiotensin I converting enzyme.

Claims (8)

A health functional food composition for preventing or ameliorating hypertension containing an arcanoid extract extracted by hydrolysis using Protamex and Neutrase as an active ingredient. delete delete delete delete The method of claim 1,
1) adding water and protamase to the demineralized crushed product of the deionized water to perform primary hydrolysis, and then deactivating the protamex;
2) dehydrogenating the protease by inactivating the protease by inactivating the protease by adding the protease to the hydrolyzate, and then deactivating the protease;
3) filtering the hydrolyzate inactivated by the Ntrease of step 2), adding ethanol and centrifuging to obtain supernatant; And
4) concentrating the supernatant of step 4) under reduced pressure, and freeze-drying the supernatant; and hydrolyzing the supernatant of step 4). The health functional food composition according to claim 1, wherein the ariposid extract inhibits the activity of angiotensin converting enzyme (ACE). A pharmaceutical composition for preventing or treating hypertension comprising, as an active ingredient, an arcanoid extract extracted by hydrolysis using Protamex and Neutrase.

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