KR20190068913A - Composition for improving inhibition activity against angiotensin converting enzyme - Google Patents

Abstract

The present invention relates to an angiotensin inhibitor having an inhibitory activity against angiotensin converting enzyme. Specifically, the present invention relates to the angiotensin inhibitor which can be usefully used for preventing or treating hypertension, kidney disease or complications thereof by containing a Dendropanax morbiferus extract, in which an angiotensin converting enzyme inhibitory activity is significantly increased by hydrolyzed with protease, trypsin, bromelain, and papain, which are hydrolases, as an active component.

Description

Technical Field [0001] The present invention relates to a composition for enhancing angiotensin converting enzyme inhibitory activity,

The present invention relates to a composition having an inhibitory activity against angiotensin converting enzyme, and more particularly, to a composition having an angiotensin converting enzyme inhibitory activity, which comprises an extract of Aspergillus oryzae Treated with a hydrolytic enzyme as an active ingredient.

Recently, nutritional status has been dramatically improved due to economic growth, improvement of education level, and scientific development, and diseases caused by changes in various lifestyle patterns including eating habits and life expectancy have also been diversified.

Cerebrovascular disease, which is the highest cause of death in Korea after cancer, is caused by hypertension and hypertension. In particular, hypertension is a leading cause of complications such as cerebrovascular disease and cardiovascular disease, and the trend is increasing as the diet is westernized and aged. Therapeutic agents such as diuretics, vasodilators, and calcium blockers are mainly used for the treatment of hypertension, but there are problems such as headache, dizziness, orthostatic hypotension, and renal dysfunction.

NO is known to relax blood vessel smooth muscle to expand blood vessels to prevent hypertension by controlling blood pressure, prevent thrombus formation that causes stroke and heart attack, and reduce arterial sclerosis by softening blood vessels. This NO is generated by decomposition of L-arginine by NOS synthase. Gamma-Amino Butyric Acid (GABA) is a non-protein amino acid that is a neurotransmitter of mammals. It has been reported that improving blood flow to the brain and inhibiting the secretion of antidiuretic hormone by acting on the blood vessel center and dilating the blood vessels to lower the blood vessels prevents hypertension.

On the other hand, the number of patients with renal insufficiency has also been gradually increasing. Such factors include changes in the living environment, an increase in the number of diabetic nephropathy due to aging or increased diabetic patients. The number of patients who are suffering from kidney failure due to renal dysfunction or the introduction of dialysis is increasing every year. In case of dialysis therapy, side effects such as erythrocyte production disorder, complications due to accumulation of microglue accumulation and increase of cardiovascular lesion are problematic . In patients with renal failure, administration of drugs that inhibit the rise of electrolytes in the blood, prescription of low-stage vaccination, and administration of erythropoietin in the case of renal anemia are performed in addition to the coercive therapy, It is thought that it is not enough to stop. In kidney diseases such as nephritis, diabetic nephropathy and renal failure, hypertension is often accompanied by hypertension, and hypertension is considered to be one of the factors causing aggravation of renal disease. Thus, depressants which are expected to inhibit the progression of renal diseases are also administered ought. Among them, the renin-angiotensin system inhibitors are particularly attracting attention.

The renin-angiotensin-aldosterone system is an endocrine system that regulates blood pressure and the volume of extracellular fluid. Renin activated by adjacent glomerular cells and secreted into the blood converts angiotensinogen to angiotensin I, and angiotensin I is converted to angiotensin II by angiotensin converting enzyme (ACE). ACE hydrolyzes the inactive C-terminal dipeptide (his-Leu) of angiotensin I and converts it to angiotensin II. Angiotensin II contracts smooth muscle smooth muscle and aldosterone secretion in the adrenal cortex Promoting increases the resorption of sodium and water in the kidneys. This increases the salinity and water content in the body and increases the total body fluid and blood pressure. In addition, the bradykinin nanopeptide having vasodilatory action is degraded to inactivate the bradykinin nanopeptide. ACE activity is a cause of hypertension, arteriosclerosis, heart failure, diabetes and various kidney diseases.

ACE inhibitors reduce the formation of angiotensin II by inhibiting the conversion of angiotensin I to angiotensin II by acting on the ACE enzyme. ACE inhibitors have been approved as therapeutic agents for the treatment of hypertension for the first time and have been known to be effective for other cardiovascular and renal diseases in the future and are being applied to various diseases such as acute myocardial infarction, heart failure, and diabetic nephropathy . Specifically, the ACE inhibitor inhibits the secretion of aldosterone, an adrenocorticotropic hormone, thereby showing a therapeutic effect on renal diseases such as renal failure, lowering blood vessel resistance in the kidney, inhibiting sodium reabsorption, It is known that diuretic action can be assisted. ACE inhibitors are believed to have a kidney-protective action that is not dependent on coercion. For example, the researchers found that the inhibitory effect of ivesartan on the progression of diabetic nephropathy was superior to control of blood pressure by calcium antagonists have. Therefore, ACE inhibitors are widely used, especially in diabetic nephropathy patients, even in the normal range of blood pressure.

In addition, the ACE inhibitor inhibits the degradation of bradykinin, exhibits vasodilatory effect, inhibits sympathetic nervous system, inhibits β1 receptor activity, and can exhibit therapeutic effect on coronary artery disease such as heart failure. In addition, it can inhibit the degradation of prostaglandins and reduce the elevated intraocular pressure due to glaucoma, and can be used to prevent visual acuity deterioration. Captopril is known as a representative ACE inhibitory drug preparation.

Synthetic ACE inhibitors are generally considered to be unsafe, and there is a growing demand for their potential for development into pharmaceuticals and foodstuffs by developing ACE inhibitors from natural products with low side effects. Known natural ACE inhibitors include cyanine hydrolyzate (Kohmura, M, et al., Agric. Biol. Chem., 54: 835 (1990)), mualeryl produced by Nocardia orientales (see Tadahiko, H. and Ryoichi. K., Biochem. Biophys. Res. (1986)), peptides obtained from porcine serum (see Karen B. et al., The Journal of Antibiotics, 37: 57: 695 (1993)), peptides of tuna intrinsic digestive enzymes (see Nobuyasu, M. and Toshio, S., Biophys. Res. Com., 139: 52 (1986) Biosci., Biotec. Biochem., 57: 922 (1933)), peptides in anchovy salted fish (refer to Matsui, T. et al., Biosci. Biotec. Biochem. 26: 321 (1993)), peptides isolated from the culture of Steptomyces species (see Uasuji, K. et al., The Journal of Antibiotics (Wakako T. et al., Nippon Shokuhin Kogyo Gakkaishi, 39: 188 (1002)) prepared from un-disaggregated rice, poly (Yoshijara, M. et al., Agric. Biol. Chem., 49: 909 (1985)), the phenolics (see contrast medium and Choi, ), And sake and its by-products (see: Nishitani Kogei, Bai Saiseisu and Isodasutori, 51: 463 (1993)). Such ACE inhibitors may be used as functional foods or drugs such as blood pressure reduction.

Crude drugs are a kind of medicines, which are used as raw materials for medicines or used as raw materials. It is known that unlike drugs made from single-component compounds, many kinds of ingredients are mixed and the effects are complex and have relatively few side effects. Dendropanax morbifera Lev . ) Is an evergreen broad-leaved arboreous tree native to the southern coast of Korea and Jeju-do. It is a species that does not fall into winter when it blooms in the winter. If it bruises the bark, it is called yellowish resin. Huangchil has been used as a rare paint for the emperor's armor, helmet, and other metal ornaments since the Three Kingdoms period. It has been used in the Goryeo period, which was written in the Goryeo period, the Guilin variety in China, And it is said that it was a special product of Baekje which was before it, and it remains in the passage of the book department of the Tang history book. It is said that it is effective for burning removal, eye quality, burn treatment, leprosy, harmlessness, and it is used recently for tea and food.

KR 100204112 B1

 Manjusri, D. and Richard, L. S., J. Biol. Chem., 250: 6162 (1975); Davis, J.O., fed. Proc., 36: 1753 (1977); Hall, J. E. et al., J. Hypertens., 4: 387 (1986)  N Engl J Med 2001 Sep 20; 345 (12): 851-60

It is an object of the present invention to provide an angiotensin converting enzyme inhibitor which comprises hydrolyzed Hwangcholgak extract as an active ingredient.

According to an aspect of the invention, the protease, trypsin, bromelain and papain hydrolysis treatment with hwangchil tree (Dendropanax Angiotensin converting enzyme (ACE) inhibitor is provided for the prevention or treatment of hypertension, kidney disease or complications thereof, which comprises an extract of Morbifera as an active ingredient.

In addition, the hydrolysis treatment may include a first step of immersing the Horticultural Raw materials in water and treating the protease and trypsin; And a second step of treating bromelain and papain in the immersing method of the whitneywood prepared in the first step.

In addition, the extract of Huacarmin can be a hot-water extract.

In addition, the hydrolysis-treated Hwigae extract can increase L-arginine and GABA content.

In addition, the complication may be caused by a complication such as arteriosclerosis, coronary artery disease, myocardial infarction, heart failure, peripheral circulation disorder, vascular stenosis, stroke, cerebral infarction, cerebral hemorrhage, ischemic brain disease, renal failure, diabetes, diabetic nephropathy, diabetic retinopathy, Edema, visual disturbances due to vascular stenosis of the eye, and glaucoma.

The angiotensin converting enzyme inhibitor according to one aspect of the present invention can exhibit significantly increased angiotensin converting enzyme inhibitory activity by hydrolysis and treatment of the extract of Hwangchujanggi as an active ingredient. Accordingly, the composition can be effectively used as a composition for preventing, ameliorating, or treating hypertension, renal disease or complications thereof acting on the renin-angiotensin-aldosterone system.

In addition, the L-arginine and GABA components are remarkably increased, and can exhibit an excellent vasodilating effect on contracted blood vessels. Through this, it can be used as a pharmaceutical composition for prevention or treatment of hypertension.

FIG. 1 shows the inhibitory activity of angiotensin converting enzyme inhibitory activity of the extract of Hwangchujang and hydrolyzed Hwangchujang extract.
FIGS. 2 and 3 show the blood vessel relaxation effect of the extract of Hwangchil-myeon and the hydrolyzed Hwacheon-myeon tree by the experiment of the base tension.

The present invention relates to an angiotensin converting enzyme inhibitor comprising Dendropanax Morbifera extract hydrolyzed with protease, trypsin, bromelain and papain as an active ingredient.

Hereinafter, the present invention will be described in more detail. In the following description, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

The angiotensin converting enzyme inhibitor of the present invention contains hydrolyzed Hwangchujang extract as an active ingredient.

The raw material of the Hwigae-jinja which can isolate the Hwigae-jinja extract may be at least one selected from the group consisting of leaves, branches, roots, and fruits of Huangchu, but is not limited thereto. It is preferred that leaves or branches of Hwangryeok tree are selected. It is more preferable that the leaves or branches of Hwigae-gil are selected. The young branch means branches of one year old or younger. This can be cut, dried and pulverized by a conventional method. The drying method is not particularly limited, and a usual method can be used. For example, natural drying, hot air drying or freeze-drying can be used.

The hydrous hydrogenated woodchuck extract can be prepared by treating hydrolysis enzymes and extracting water with a solvent. In the production of the hydrolyzed Hokutogi extract of the present invention, the preferred sequence of the embodiment is to immerse the raw material in water, hydrolyze the raw material in two steps, heat it to extract, and perform enzyme inactivation have. Wherein the hydrolysis treatment comprises: a first step of immersing the Horticulte raw material in water and treating the protease and trypsin for primary hydrolysis; And a second step of treating the bromelain and papain for hydrolysis in the immersing method of the whitish wood prepared in the first step. In the present invention, the hydrolysis treatment of the raw material is carried out in two stages as described above, so that the enzymes having different reaction temperature conditions can be reacted at the optimal temperature, while the optimal reaction temperatures are similar, The enzymatic groups can be treated together at the same stage to shorten the reaction time, thereby shortening the time required for the whole process and increasing the productivity. The enzyme to be added during the hydrolysis treatment may be treated in the form of an enzyme solution dissolved in a solvent.

In the first step of the hydrolysis treatment, the crude Horticulture raw material is first immersed in water and subjected to primary hydrolysis treatment by adding a proteolytic enzyme for primary hydrolysis treatment. Specifically, the first step of the hydrolysis treatment may be performed by adding a protease and trypsin to the immersing solution of Hwangchil, which is the water immersed in the Hwangchu tree. The water forms a reaction pool for immersion solvent and enzyme reaction, and can be used in a ratio of 7 to 15 times the weight of the raw material of Horticultural wood. The protease may be added to increase the amount of protein eluted from the Hwangchol-yaw raw material, and the protease derived from Bacillus subtilis is preferably selected. The trypsin may be added to break down the Hwangchil-derived protein into a polypeptide and a small number of amino acids. The protease and trypsin to be treated in the first step may be treated in an amount of 0.1 to 1% by weight based on 100% by weight of the total of the raw materials of Horticulture. The first step may be carried out at 40 to 50 < 0 > C for 5 to 15 hours. Preferably at 42 to 47 DEG C for 6 to 10 hours.

Next, the second step of the hydrolysis treatment can be carried out. The second step may be carried out by treating the amino acid-digesting enzyme of the crude Horticulturae which has been treated with the proteolytic enzyme in the previous step. In detail, the treatment can be performed by adding bromelain and papain to the protease and trypsin-treated Yellowish tree immersion liquid prepared in the first step. The bromelain and papain treated in the second step may be treated in an amount of 0.1 to 1 wt% based on 100 wt% of the sulfur-free raw material. The first step may be carried out at 50 to 60 < 0 > C for 12 to 20 hours. Preferably, the treatment is carried out at 53 to 57 캜 for 14 to 18 hours. In the second step, bromelain and papain may be prepared and treated with an enzyme solution dissolved in a solvent, and the solvent used for preparing the enzyme solution may be 0.5 to 2 times as much as the weight of the raw material of Hokutogi, but is not limited thereto.

Next, hot water extraction may be performed on the immersed Hwigulak tree immersion liquid by the method including the first and second steps, in order to inactivate the treated enzyme and separate the Hwigulnghi extract as a useful ingredient . Through this process, it is possible to increase the productivity by decreasing the total manufacturing time and effort while improving the convenience in the production process, by making the extract of Hwangchujang and inactivating the enzyme by one process. The treatment temperature at the hot water extraction may be 80 to 120 캜, preferably 100 to 120 캜. Extraction for 1 to 72 hours within the above-mentioned temperature range may be carried out to isolate and extract the extract of Hokutogi (Mushrooms) and enzyme inactivation. Preferably 2 to 12 hours, more preferably 3 to 7 hours. If necessary, water may be further extracted by adding water in an amount of 8 to 12 times as much as the weight of the raw material of horticultural to increase the extraction yield. The hydrolyzed Hwangchu tree extract prepared through hydrothermal extraction can be provided in the form of a concentrated liquid or solid powder by controlling the concentration by removing the solvent through filtration, concentration or drying.

The hydrolysis-treated Hwilinchu extract of the present invention has excellent angiotensin converting enzyme inhibitory activity. In detail, the angiotensin converting enzyme inhibiting activity of the crude oilwood raw material can be enhanced by hydrolyzing the crude oil wood raw material. Thus, the inhibition of angiotensin I to angiotensin II conversion is effective in reducing the increase in body salinization, increase in total body fluid and blood pressure caused by the action of angiotensin II, and prevention of hypertension, kidney disease or complications thereof , ≪ / RTI > For example, it can be used for the improvement or treatment of hypertension, kidney disease or complications thereof as a diuretic agent which promotes the discharge of body fluids. Such complications include, for example, atherosclerosis, coronary artery disease, myocardial infarction, heart failure, peripheral blood circulation disorder, vascular stenosis, stroke, cerebral infarction, cerebral hemorrhage, ischemic brain disease, renal failure, diabetes, diabetic nephropathy, diabetic retinopathy , Proteinuria, edema, visual disturbances due to vascular constriction of the eyes, and glaucoma. In addition, the hydrolysis-treated Hwigulak tree extract of the present invention can remarkably amplify the content of L-arginine and GABA, which are useful components of the raw material of Hwangjak wood, through the hydrolysis treatment at the same time, Effect can be shown. The hydrolyzed Hexacanthus japonica extract of the present invention has excellent angiotensin converting enzyme inhibitory activity and has an increased L-arginine and GABA content. Therefore, the present invention relates to a method for preventing, ameliorating, or treating vascular disorders associated with vasoconstriction Can be used as a composition of high quality and high efficiency. Examples of the vascular disorder diseases include hypertension, arteriosclerosis, coronary artery disease, myocardial infarction, heart failure, (peripheral) blood circulation disorder, vascular stenosis, stroke, cerebral infarction, cerebral hemorrhage or ischemic brain disease.

The angiotensin converting enzyme inhibitor containing the hydrolyzed Hwangchu-myeon extract of the present invention as an active ingredient may be prepared as a pharmaceutical preparation for use in the prevention or treatment of hypertension, kidney disease or complications thereof. The pharmaceutical preparations of the present invention can be prepared by using at least one selected from the group consisting of pharmaceutically acceptable carriers, diluents and excipients according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by formulating it into a unit dose form or by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules. The formulation may further comprise a dispersant or stabilizer.

If desired, the pharmaceutical preparation may further comprise a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are those conventionally used in the field of the present invention and include for example lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, But are not limited to, crystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

If desired, the pharmaceutical preparations of the present invention may further contain additives known in the art in addition to the above components. For example, it may further include at least one selected from the group consisting of a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, and a preservative agent, but is not limited thereto.

The pharmaceutical preparation of the present invention can be administered orally or parenterally. In case of parenteral administration, it can be administered by skin application, rectal injection, intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection and transdermal administration. A suitable dosage of the pharmaceutical composition of the present invention may be controlled by such factors as the formulation method, the manner of administration, the age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient And the ordinarily skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. The daily dosage of the pharmaceutical composition of the present invention may be 0.001 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg. If necessary, the above dose may be divided into several times and administered at predetermined time intervals. The daily dose of the pharmaceutical composition of the present invention can be adjusted by a skilled physician.

The angiotensin converting enzyme inhibitor containing the hydrolyzed Hwigae-jinja extract of the present invention as an active ingredient can be produced as a functional food. The functional food is a food prepared by adding a hydrolyzed Hwangcholgak extract to a food material, or a food prepared by using the extract as a capsule, a powder, or a suspension. In the case of ingestion, Food. Examples of the food include drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snack, confection, pizza, noodle, gum, ice cream, soup, beverage, tea, vitamin complex But is not limited thereto.

If necessary, the functional food of the present invention may further comprise components that are ordinarily added in food production. For example, there may be mentioned nutrients, vitamins, minerals (electrolytes), flavors, coloring agents, thickening agents, pectic acid, pectate salts, alginic acid, alginates, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, , Carbonating agent, pulp, and the like, but is not limited thereto.

In one embodiment of the present invention, the functional food may be a drink. The drink may further contain flavors or natural carbohydrates. The natural carbohydrate may be monosaccharide (e.g., glucose, fructose, etc.); Disaccharides (e.g., maltose, sucrose, etc.); oligosaccharide; Polysaccharides (e.g., dextrin, cyclodextrin and the like); And sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.). Examples of the flavoring agent include natural flavoring agents (e.g., tau Martin and stevia extract) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.), alone or in combination.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail with reference to the following examples. It will be apparent to those skilled in the art that these embodiments are illustrative of the present invention and are not intended to limit the scope of the appended claims and that various changes and modifications may be made to the embodiments within the scope and spirit of the invention , And it is natural that such variations and modifications fall within the scope of the appended claims.

Example

≪ Preparation of an extract of Wuchulia chinensis>

Leaf to the green part within 50cm from the end of the leaves was cut into 2㎝ of raw material and hot air drying was carried out. Twenty times as much distilled water as the weight of the raw material was added to the leaves, and the mixture was subjected to hot water extraction at 100 ° C for 4 hours, then concentrated to a concentration of 30brix or more and lyophilized to obtain a powdery hot water extract. Using a raw material of 2.5 kg, 521.55 g of the extract was obtained and the yield was found to be 20.9% (w / w).

≪ Preparation of hydrolyzed Hwangchulchae extract >

One kilogram of dried yellowtail leaves were used as raw materials, and distilled water in an amount of 9 times as much as the raw material weight was added to the raw materials and immersed. 1 g of protease and 1 g of trypsin were dissolved in distilled water and subjected to simultaneous immersion extraction and primary hydrolysis at 45 ° C for 8 hours. After the completion of the first hydrolysis, 1 g of bromelain and 1 g of papain were dissolved in distilled water in an amount of 1: 1 based on the weight of the raw material, and the solution was treated with the immersion liquid and subjected to secondary hydrolysis at 55 ° C for 16 hours . After the completion of the secondary hydrolysis, distilled water in an amount of 10 times as much as the weight of the raw material was further added, and the mixture was subjected to hot water extraction at 100 DEG C for 4 hours in a state of immersing the raw material of Horticornia in distilled water in a total amount of 20 times as much as the weight of the raw material, And the process of doing it. After that, the solution was concentrated to a concentration of 30 brix or more and lyophilized to obtain a hot-water extract of powder form.

<Determination of L-arginine and GABA content of Wuchu tree extract>

The contents of L-arginine and GABA were analyzed on the extracts of Hwangchil-myeon and hydrolyzed Hwangchu-myeon, and hydrolysis treatment was carried out to confirm the amplification rate of useful components. As a comparative example, the hydrolysis-treated Hwigulnyeo extract of the present invention was used as an example. A 6410A triple quadrupole mass spectroscopy (Agilent, USA) coupled with a 1200 series LC (Agilent, USA) was used for the analysis of L-arginine and GABA content. The nebulizer pressure, N ₂ gas flow rate and temperature were 40 psi, 10 L / min, and 310 ° C, respectively, in a multiple reaction monitoring (MRM) mode after the sample was ionized with a positive electro-spray ionization And the capillary voltage was maintained at 4 kV. Gemini-NX C18 (4.6 × 150 mm, 3 μm, Phenomenex) was used as the column. The sample injection volume was 5 μl and the column oven was kept at 35 ° C. Starting with 0% (v / v) B and 95% (v / v) using water (A) and methanol (B) mixed with 5 mM ammonium acetate and 0.1% (v / v) formic acid, B to 0% (v / v) B and analyzed for 30 minutes. The flow rate was maintained at 0.5 ml / min. Analysis for each sample was repeated two times and the results are shown in Table 1 below. In the following Table 1, the L-arginine and the GABA content units are mg / g.

division Measure
repeat L-arginine
(Mg / g) Gabba
(Mg / g) Comparative Example

1 time 3.45 19.975 Episode 2 2.903 18.017 Average 3.177 18.996 Example

1 time 16.840 25.176 Episode 2 18.730 30.450 Average 17.770 27.813

As shown in Table 1, it can be seen that the L-arginine and the GABA content of the hydrolyzed Hwangchu-wood extract were increased compared to the Hwangchu-wood extract. Based on the comparative example, the L-arginine content of the example was found to be 559% (w / w) and the GABA content was found to be 146% (w / w).

<ACE inhibition activity confirmation>

Angiotensin converting enzyme (ACE) inhibitory activity was measured by the spectrophotometric assay and the properties of the angiotensin-I converting enzyme of rabbit lung, Biochem. Pharmacol., 20, 1637-1648 described by Cushman and Cheung (1971) Some modifications were made. Hippuric acid-Histidine-Leucine (Hip-His-Leu, Sigma Chemical Co., St. Louis, USA) was used as a substrate for measuring ACE inhibitory activity. Hippuric acid, The results of this study are as follows. For analysis, 100 μL (pH 8.3) of a 0.4 M sodium borate buffer containing 0.3 M sodium chloride (NaCl) was added to the extracts of the comparative example or the example at concentrations of 0.03, 0.1, 0.3, 1, 3 and 10 μg / 50 μL and 0.3 μL of sodium borate buffer solution containing 0.3 M NaCl were mixed with 100 μL of 5 mM Hip-His-Leu substrate solution and incubated at 37 ° C. for 30 minutes. Then, 100 mU / mL angiotensin converting enzyme 50 占 퐇 of the enzyme solution was added and reacted at 37 占 폚 for 1 hour. After the reaction was completed, 100 μL of 1 M HCl was added to stop the reaction. To extract the hexuronic acid, 1 mL of ethylacetate was mixed and sufficiently mixed with a vortex mixer. After centrifugation at 3000 × g for 15 minutes, 500 μL of the supernatant was completely dried in a dryer at 65 ° C., 1 mL of 1 M sodium chloride was added to dissolve, and the absorbance was measured at a wavelength of 228 nm using a microplate spectrophotometer. At this time, the content of the sample extract required to represent 50% of the ACE inhibitory activity was expressed by IC ₅₀ value. The ACE inhibitory activity of each of the comparative examples and the examples was quantitatively analyzed, and the inhibitory effect thereof was shown in Fig. 1 and Table 2 in terms of% of the untreated group. The non-treated group was prepared by adding distilled water instead of the extract of the comparative example or the example. In Fig. 1, (A) is the measurement result of the comparative example and (B) is the measurement result of the embodiment. Measurements were performed 4 times each. Captopril, known as ACE inhibitor, was used as a control for the comparison of ACE inhibitory activity. The inhibitory effect of captopril as a control group, IC _50, was 9.47 μM.

Condition density
(μg / ml) Repeat measurement Inhibitory effect
(%) Repeat measurement Inhibitory effect
(%) Repeat measurement Inhibitory effect
(%) Repeat measurement Inhibitory effect
(%) Comparative Example 0.03 1st round -3.00 Second round 0.00 Three times 6.00 Four times 8.00 0.1 1st round -4.00 Second round 1.25 Three times 7.25 Four times 2.00 0.3 1st round -0.86 Second round -0.86 Three times 1.29 Four times 2.14 One 1st round 0.69 Second round 0.95 Three times 0.95 Four times 1.00 3 1st round 4.00 Second round 4.75 Three times 7.75 Four times 7.75 10 1st round 10.75 Second round 13.00 Three times 7.00 Four times 11.00 30 1st round 10.75 Second round 13.00 Three times 11.00 Four times 13.00 100 1st round 22.00 Second round 25.00 Three times 31.00 Four times 30.00 300 1st round 44.00 Second round 46.00 Three times 46.00 Four times 52.00 Example 0.03 1st round 3.95 Second round 4.25 Three times 4.40 Four times 4.40 0.1 1st round 8.00 Second round 8.20 Three times 9.20 Four times 9.30 0.3 1st round 11.38 Second round 11.75 Three times 12.38 Four times 14.25 One 1st round 12.38 Second round 13.50 Three times 14.25 Four times 13.88 3 1st round 15.32 Second round 15.00 Three times 14.68 Four times 14.53 10 1st round 25.75 Second round 25.00 Three times 25.00 Four times 23.75 30 1st round 32.33 Second round 32.00 Three times 31.33 Four times 30.00 100 1st round 37.00 Second round 38.00 Three times 40.00 Four times 45.00 300 1st round 63.33 Second round 73.33 Three times 75.00 Four times 75.00

1 and Table 2, in the case of the comparative example, the inhibition effect was as low as 47% on the average compared with the untreated group when treated at the concentration of 300 μg / ml, And 71.7% respectively. As a result, it was confirmed that the hydrolyzed Hwangchu-myeon extract of the present invention showed significantly increased ACE inhibitory activity compared to the hydrolysis-treated Hwangchu-myeon extract. The hydrolysis-treated Hwigulak tree extract of the present invention is excellent in ACE inhibitory activity and can easily inhibit the conversion of angiotensin I to angiotensin II, and thus can be applied to hypertension, kidney disease or complications thereof to prevent, Can be usefully used as a composition for treatment.

<Assessment of blood pressure lowering effect following thoracic aortic vessel relaxation>

Vascular tissue sections of rats were prepared for the experiment. Sprague-Dawley rats (Samtako, Korea) were weighed and placed in a feeding room with a solid feed (LabDiet 5L79, Orient Bio, Korea) and water at 160 to 200g Respectively. The rat was stunned and immediately bled. The rat was then opened along the midline of the abdomen, and the thoracic aorta was separated and cut into a ring shape with a length of about 2 mm. An oxygen saturated krebs solution (Krebs buffer (mM / L); 118.4 NaCl, 25 NaHCO ₃ , 11.66 glucose, 4.7 KCl, 1.18 MgSO ₄ .7H ₂ O, 2.5 CaCl ₂ .2H ₂ O, 1.19 KH ₂ PO ₄ , pH 7.4). While the oxygen was continuously supplied, the surrounding adipose tissue and connective tissue were cleanly removed, and then cut into transverse sections of about 2 mm in length to form a ring section.

To investigate the effects of the extracts of Hwangchulchu and hydrolyzed Hwangchu - wood extracts on the relaxation capacity of rat thoracic aortic vascular tissues, we first measured the functional activity of vascular aortic vascular tissues in the rat. The solution in the organ chamber maintained aeration at 95% oxygen (O ₂ ) -5% carbon dioxide (CO ₂ ) mixture at 37 ° C, pH 7.4. Tension of the blood vessels was measured using a polygraph system (PowerLab 8/35, AD Instruments, Dunedin, New Zealand) connected to an isometric transducer (MLT0201, Panlab, Cornella, Spain) and a computer analyzer Pro V8-MLS260 / 8, ADInstruments, Dunedin, New Zealand). Before the drug experiment, thoracic aortic blood vessels underwent baseline tension by applying one gram of force to a blood vessel section with one side of the vascular tissue hanging on a ring placed under the fascia and the other side suspended on a force-displacement transducer. After keeping the equilibrium, if the baseline was maintained, Phenylephrine 10 ^-5 M was administered, and the acetylcholine 10 ^{- 5} M, 10 ^{- 4} M, 10 ^{- When 3} M was added, complete relaxation was measured. In addition, when Phenylephrine 10 ^-5 M was shrunk and exposed to a Krabs solution, it was confirmed that the reaction was stable and the functional activity could be determined. This is shown in Fig. 2 (A).

After confirming the functional activity of the vascular tissue of the rat thoracic aorta, experiments were carried out using comparative examples and examples. Phenyl LES principal (Phenylephrine) 10 ^{- 5} administered in M concentration by the after-induced vasoconstriction, comparative example general hwangchil wood extracts 0.1, 0.3, 1, the relaxation degree of the contracted blood vessel and administered in 3, 10μg / ml And the results are shown in Fig. 2 (B). In order to analyze the effects of the hydrolyzed Hwangchu-myeon extract, which is an embodiment, on the rat thoracic aortic vessel relaxation response, 0.1, 0.3, 1, 3, and 10 μg / ml And the results are shown in Fig. 2 (C). The results of the comparison of the relaxation effects by the treatment concentrations of the comparative example and the example are shown in Fig.

Referring to Figures 2 and 3, the EC ₅₀ value of the comparative example was 6.61 ± 0.06 μg / ml, and the EC ₅₀ value of the example was 3.50 ± 0.07 μg / ml. It can be confirmed that the blood vessel relaxation effect of the example is twice as excellent as that of the comparative example. Comparing the IC ₅₀ values, the comparative example is 6.61 μg / ml and the example is 3.50 μg / ml. The results show that the ACE inhibitor containing the hydrolyzed Horticulti extract of the present invention , It can be seen that the same effect can be obtained even when the amount of intake corresponding to the half of the extract obtained from the extract obtained from the simple extract by the general method is used.

The ACE inhibitor of the present invention enhances ACE inhibitory activity through hydrolytic enzymatic treatment and further contains vinegar extract as an active ingredient in which L-arginine and GABA are simultaneously amplified, thereby preventing or treating hypertension. And thus can be effectively applied as a highly effective therapeutic agent.

Claims (5)

Containing an extract of Dendropanax Morbifera hydrolyzed with protease, trypsin, bromelain and papain as an active ingredient,
Angiotensin converting enzyme (ACE) inhibitor for the prevention or treatment of hypertension, kidney disease or complications thereof.
The method according to claim 1, wherein the hydrolysis treatment comprises: a first step of immersing the woody raw material in water and treating the protease and trypsin; And
Wherein the second step is to treat bromelain and papain in the immersing method of the whitneywood prepared in the first step.
Angiotensin converting enzyme (ACE) inhibitor for the prevention or treatment of hypertension, kidney disease or complications thereof.
The method according to claim 1, wherein the extract is a hot-water extract.
Angiotensin converting enzyme (ACE) inhibitor for the prevention or treatment of hypertension, kidney disease or complications thereof.
2. The method according to claim 1, wherein the hydrolyzed Hexachrichum extract has increased L-arginine and &lt; RTI ID = 0.0 &gt;
Angiotensin converting enzyme (ACE) inhibitor for the prevention or treatment of hypertension, kidney disease or complications thereof.
The method of claim 1, wherein the complication is selected from the group consisting of atherosclerosis, coronary artery disease, myocardial infarction, heart failure, peripheral blood circulation disorder, vascular stenosis, cerebral ischemia, cerebral hemorrhage, ischemic brain disease, renal failure, diabetes, diabetic nephropathy, Characterized in that it is a symptom or a disease selected from the group consisting of acute corneal edema,
Angiotensin converting enzyme (ACE) inhibitor for the prevention or treatment of hypertension, kidney disease or complications thereof.

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