KR102151738B1 - Composition for Anti-hypertension Using Protamex Hydrolysates of Hippocampus abdominalis, Fractions Thereof, or Effective Peptides Thereof - Google Patents

Abstract

Disclosed is an antihypertensive composition using a Hippocampus abdominalis hydrolyzate, fraction thereof, or effective peptide thereof.

Description

Composition for Anti-hypertension Using Protamex Hydrolysates of Hippocampus abdominalis, Fractions Thereof, or Effective Peptides Thereof}

The present invention relates to a composition for antihypertension using a Big Valley hippocampus protamex hydrolyzate, a fraction thereof, or an effective peptide thereof.

Recently, as interest in aging and adult diseases has risen, studies to prevent diseases by eliminating excessively generated reactive oxygen in the living body have been actively conducted. Reactive oxygen is an electron acceptor of oxygen, which is absolutely necessary for the maintenance of life in aerobic organisms, and occurs in the process of continuing biochemical reactions to supply energy. 2-3% of the oxygen introduced into the body is is converted into active oxygen (Reactive Oxigen Species) by the electron transport processes and energy metabolism, and a variety of external stimuli in ^{_{cells, superoxide radical (O 2 -)}} , hydroxyl radical ( and OH) and hydrogen peroxide (H ₂ O ₂ ), a non-free species.

Such ROS not only oxidizes lipids constituting cell membranes, but also damages DNA and affects mutagenesis, causing chronic degenerative diseases such as arteriosclerosis, cancer, hypertension, and diabetes (IJBCB. 2007, 39(1):44~) 84; J. Hepatology. 2007, 46(2):193-197). Reports that ROS is an important factor involved in cardiovascular diseases including hypertension indicates a close relationship between free radical oxidation and hypertension (American J. of hypertension. 2004, 17(9):809~816; Free radical biology & medicine) 2006, 40:2028~2039).

Hypertension refers to the case where the average blood pressure measured several times is 140mmHg or more in systolic blood pressure and 90mmHg or more in diastolic blood pressure, and it is emerging as an important problem for modern people's health by artificially increasing blood pressure during human physiological activities. An abnormal increase in blood pressure due to various causes rather than caused by one reason is called primary hypertension or essential hypertension. In addition, high blood pressure may occur due to the surrounding environment, but blood pressure may rise due to genetic causes, which is called secondary hypertension.

On the other hand, angiotensin converting enzyme (ACE), an important enzyme in the renin-angiotensin system (RAS), an important mechanism of increasing blood pressure, is an enzyme that converts angiotensin Ⅰ (AngI) to angiotension Ⅱ (AngII). AngII converted into AngII type 1 It is known to act on the (AT1) receptor to constrict blood vessels and increase blood pressure by increasing the secretion of aldosterone. In addition, AngII increases the production of reactive oxygen species and nitrogen species by activating the NADH/NADPH oxidase system, inducing oxidative stress in endothelial cells and inducing inflammatory reactions in blood vessels through oxidation of low-density lipoproteins (Am J. Cardiol). 2003, 91(12A)).

The currently used angiotensin converting enzyme inhibitors are synthetic chemicals or synthetic peptides. Synthetic chemicals and synthetic peptides are useful in that they can be produced by mass synthesis, but itching, headache, and tachycardia , Chest pain, weakness, and other side effects have been reported. Accordingly, there is an urgent need for new angiotensin converting enzyme inhibitors without side effects.

The hippocampus is used worldwide as a drug to treat asthma, heart disease, fractures, etc. In particular, Hippocampus abdominalis is the largest among seahorses and has a beautiful body color and body shape, so it is popular in the international seawater coronary life market due to its high coronal value. There are many. The head is at a right angle to the body, the tail can be wrapped around the substrate and attached, and the male has the habit of protecting the egg. It is used as a medicinal herb in Asia and elsewhere, and is a very popular fish because of its unique shape (Technical report series, 2003, 4:15).

However, due to its medicinal value, some countries are overfishing, and furthermore, the number of natural entities is gradually decreasing as the environment that can inhabit due to global warming and environmental pollution on the coast is reduced. In Australia and neighboring countries, China and Korea, artificial insemination is being used, but the number is significantly insufficient compared to demand (Aquaculture. 2000, 190:377-388). Immune response and physiological studies are being conducted in the hippocampus ( Hippocampus kuda Bleeker), hippocampus guttu-latus , and short-snouted seahorse ( Hippocampus hippocampus ).

Therefore, the present inventors confirmed that the Big Valley hippocampus ( Hippocampus abdominalis ) protamex hydrolyzate, its fraction, and a novel peptide isolated therefrom exhibit an ACE (angiotensin converting enzyme) inhibitory activity, as a new resource having antihypertensive activity. I want to use it.

An object of the present invention is to provide a composition for antihypertension using a Big Valley hippocampus protamex hydrolyzate, a fraction thereof, or an effective peptide thereof.

Other or specific objects of the present invention will be presented below.

The present inventors prepared a Big Belly hippocampus protamex hydrolyzate, three fractions according to the molecular weight of the hydrolyzate, and four gel filtration fractions of this fraction, as confirmed in the Examples and Experimental Examples below, and gel filtration. Separation of three single peptides from the fractions, and evaluation of ACE (angiotensin I converting enzyme) inhibitory activity for these fractions and peptides, it was confirmed that the hydrolyzate, the fraction, and all three single peptides had ACE inhibitory activity. .

The present invention is presented on the basis of these experimental results, in one aspect, the present invention is a Big Belly hippocampus protamex hydrolyzate, a fraction of the hydrolyzate, any one of the three peptides in Table 1 below, or of the peptide It relates to a composition for improving hypertension comprising a pharmaceutically acceptable salt as an active ingredient.

In the present specification, "Big Belly hippocampus protamex hydrolyzate" refers to a group of substances containing amino acids and/or peptides as an enzyme-treated product obtained by hydrolyzing the Big Belly hippocampus with Protamex, a kind of protease. Such an enzyme-treated product is used as it is or by taking the supernatant after centrifugation, or the enzyme-treated product or its supernatant is concentrated under reduced pressure and/or lyophilized to be used in liquid or powder form, or the enzyme-treated product as it is or the supernatant is used in water or ethanol. The extract may be extracted with an organic solvent such as, or the extract may be fractionated using an organic solvent having a different polarity, or the extract or fraction may be concentrated and used. Here, the organic solvent is a lower alcohol having 1 to 4 carbon atoms including ethanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO ), 1,3-butylene glycol, propylene glycol, and a mixed solvent thereof.

In addition, in the present specification, "the fraction of the Big Belly hippocampus protamex hydrolyzate" refers to a fraction obtained by using a Big Belly hippocampus protamix hydrolyzate or an extract of water or organic solvent of this hydrolyzate using organic solvents having different polarities , Refers to a fraction obtained by fractionation according to molecular weight or a gel filtration fraction of these fractions. The organic solvent is a lower alcohol having 1 to 4 carbon atoms including ethanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO ), 1,3-butylene glycol, propylene glycol, and a mixed solvent thereof, and the like, and the elution solvent during gel filtration may be water or the organic solvent.

In addition, in the present specification, "protamex" refers to an enzyme having endopeptidase activity that recognizes and degrades hydrophobic amino acids (Food Chem. 2012; 134:1360-1367; Austin J Nutri Food Sci. 2014; 2(10)) :1053).

In addition, in the present specification, the term "active ingredient" refers to a component capable of exhibiting a desired activity alone, or capable of exhibiting activity together with a carrier that is not itself active.

In addition, in the present specification, the term "pharmaceutically acceptable salt" refers to a salt that does not inhibit the activity of the peptide compound of the present invention, and an organic salt that is generally used for the formation of a metal salt or an acid addition salt (eg, acetate, citrate, Oleate, oxalate, glutonate, etc.) or inorganic salts (such as chloride, antioxidant, borate, carbonate, etc.) may be used.

In addition, in the present specification, "improvement" is meant to include alleviation of symptoms of a disease, treatment of a disease, and prevention of a disease.

The composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit a blood pressure lowering effect or an improvement effect of essential hypertension intended for treatment, etc., depending on the formulation, etc. It will be determined within the range of 0.001% to 15% by weight based on the total weight. Herein, "effective amount" refers to the intended medical treatment, such as the effect of lowering blood pressure or improving essential hypertension, etc., when the composition of the present invention is administered to a mammal, preferably a person, to which the composition of the present invention is administered during the administration period according to the advice of a medical professional It refers to the amount of the active ingredient contained in the composition of the present invention that can exhibit pharmacological effects. Such effective amounts can be determined empirically within the range of ordinary skill in the art.

In addition to the active ingredient, the composition of the present invention is already safe in the art for increasing or reinforcing the effect of improving hypertension, or enhancing the convenience of intake or intake through the addition of similar activities such as fatigue improvement activity and antioxidant activity. Any compound or natural extract that has been validated and known to have the corresponding activity may further be included.

These compounds or extracts include compounds or extracts, pharmaceuticals, etc. listed in the pharmacopoeia of each country ("Korean Pharmacopoeia" in Korea), health functional food code of each country ("health functional food standards and standards" notified by the Ministry of Food and Drug Safety in Korea), etc. In accordance with the laws of each country governing the manufacture and sale of products (the “Pharmaceutical Affairs Act” in Korea), laws governing the manufacture and sale of compounds, extracts, and health functional foods that have been approved for items (in Korea, the “Health Functional Food Act” ”), the functional compound or extract is included. For example, L-glutamic acid-derived GABA-containing powder, which is recognized as a'blood pressure control' in accordance with the Korean Health Functional Foods Act, Gatsuobushi oligo peptide, natto bacteria culture powder, Seomoktae peptide complex, salmon peptide, Olive leaf extract, green tea extract, bamboo leaf extract, melon extract, bokbunja extract (powder) recognized as functional as'antioxidant', fermentation-produced amino acid complex that is recognized as a'fatigue improvement', dulcis fruit extract, Honggyeongcheon Extracts and the like will correspond to these compounds or extracts.

One or more of these compounds or natural extracts may be included in the composition of the present invention together with the active ingredient.

The composition of the present invention can be grasped as a food composition in a specific aspect. When the composition of the present invention is understood as a food composition, its use may be understood as inhibiting skin hypersensitivity reactions or inhibiting hypersensitivity reactions.

The food composition of the present invention may be prepared in any form, such as beverages such as tea, juice, carbonated beverages, ion beverages, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, snacks, and noodles. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrup, gels, jelly, can be prepared in health functional food preparations such as bars.

In addition, the food composition of the present invention can be classified as a product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in terms of legal and functional classification. For example, it is a health functional food according to the Korean 「Health Functional Food Act」, or confectionery, bean, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 (``Food Standards and Standards'' notified by the Ministry of Food and Drug Safety). , Special use food, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to food and mixed or infiltrated in manufacturing, processing, or preserving food. Since they are consumed daily and for a long time with food, their safety must be ensured. In the Food Additive Code (“Food Sanitation Act” in Korea) governing the manufacture and distribution of food, food additives with guaranteed safety are limited in terms of ingredients or functions. In the Korean Food Additives Code (“Food Additive Standards and Standards” notified by the Ministry of Food and Drug Safety), food additives are classified into chemical synthetic products, natural additives and mixed preparations in terms of ingredients.These food additives are sweeteners and flavors in terms of function. It is divided into agent, preservative, emulsifier, acidulant, thickener, etc.

Sweeteners are used to impart a suitable sweetness to food, and both natural and synthetic can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to improve taste or flavor, and both natural and synthetic can be used. Preferably, it is the case of using a natural one. In the case of using natural ingredients, the purpose of nutrient enhancement can be combined in addition to flavor. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or may be obtained from green tea leaves, roundtails, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, you can use those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo. The natural flavoring agent may be a liquid concentrate or a solid extract. In some cases, synthetic flavoring agents may be used, and esters, alcohols, aldehydes, terpenes, and the like may be used as synthetic flavoring agents.

As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin, etc. may be mentioned, and as the acidulant, arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like can be used. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms in addition to the purpose of enhancing taste.

As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may include a physiologically active substance or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutritional properties and ensuring stability as a food additive.

Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoyl thiamine, etc., and minerals include calcium preparations such as calcium citrate, magnesium stearate. Magnesium preparations such as, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, and the like.

In the food composition of the present invention, the food additive as described above may be included in an appropriate amount to achieve the purpose of addition according to the product type.

With respect to other food additives that may be included in the food composition of the present invention, reference may be made to the food code of each country or the food additive code.

The composition of the present invention may be understood as a pharmaceutical composition in other specific embodiments.

The pharmaceutical composition of the present invention may be prepared in an oral dosage form or a parenteral dosage form according to an administration route by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including a local route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is a case in which two or more formulations of drugs are combined according to the route of administration, for example, when one drug is firstly administered by an intravenous route and secondly, the other drug is administered by a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopeias of each country, including the "Korean Pharmacopeia."

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to a method known in the art together with a suitable carrier It can be prepared in such a formulation. Examples of suitable carriers at this time include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, grease Serol, etc. are mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, colorants, and diluents may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferryst, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc., and oleic acid as a lubricant Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polydecylene glycol, etc., and as disintegrating agents starch, methyl cellulose , Agar, bentonite, xanthan gum, starch, alginic acid, or a sodium salt thereof. Moreover, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. are mentioned as a diluent.

When the pharmaceutical composition of the present invention is prepared in a parenteral formulation, it may be formulated in the form of an injection, a transdermal administration, a nasal inhalation and a suppository according to a method known in the art together with a suitable carrier. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, triethanol amine-containing PBS (phosphate buffered saline), sterile water for injection, or an isotonic solution such as 5% dextrose may be used. . When formulated into a transdermal dosage form, it can be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, liniment, air roll, and the like. Nasal inhalants can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide, and when formulated as a suppository, the carrier is Withepsol ( witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like.

Regarding the specific formulation of the pharmaceutical composition is known in the art, for example, Remington's Pharmaceutical Sciences (19th ed., 1995) and the like can be referred to. These documents are considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. May be in the /kg range. Administration can be made once a day or divided into several times. These dosages should not be construed as limiting the scope of the invention in any aspect.

As described above, according to the present invention, it is possible to provide a composition for antihypertension using a Big Valley hippocampus protamix hydrolyzate, a fraction, or a peptide isolated therefrom. The composition of the present invention can be commercialized as food, medicine, and the like.

1 is a schematic diagram of separation of protamex hydrolyzate of Big Valley hippocampus and fractions of the hydrolyzate according to three molecular weights.
FIG. 2 is an elution profile of four gel filtration fractions (F1 to F4) of the third fraction (MW: ≤ 5 kDa) of the fractions according to the three molecular weights of the protamex hydrolyzate of the Big Valley hippocampus.
3 to 5 are the results of sequence analysis of the three peptides by a Q-TOF ESI mass spectrometer.
6 to 8 are molecular weight analysis results of the three peptides by a Q-TOF ESI mass spectrometer.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.

<Example> Hippocampal enzyme hydrolyzate-derived peptide separation and identification

Big Belly Seahorse ( Hippocampus abdominalis ) was provided frozen from Haecheonma (Jeju), located in Gujwa-eup, Jeju City. The whole hippocampus including the intestines was freeze-dried and finely ground to take 10g. , The proteolytic enzyme Protamex (Protamex, Novozyme Co. Denmark) was treated with an enzyme-to-substrate ratio of 1:100 (w/w) and hydrolyzed for 24 hours while stirring at 100 rpm at pH 6.0 and 40°C. . Next, the hydrolyzate was neutralized and centrifuged at 10000 rpm for 10 minutes, followed by filtration to separate only the supernatant. The separated supernatant was inactivated for 10 minutes in a water bath at 100° C. and then lyophilized and used in the experiment (FIG. 1).

The lyophilized enzyme hydrolyzate was 4°C with an ultrafiltration membrane with a molecular weight cut-off (MWCO) of 5 kDa and 10 kDa, respectively, using a Millipore's Lab scale TFF system (Millipore Corporation, Bedford, Massachusetts, USA). Fractionation was performed in to obtain three fractions (MW: ≥10 kDa, MW: 5-10 kDa, MW: ≤5 kDa) (FIG. 1).

For the three fractions, the ACE inhibitory activity was evaluated as in the experimental examples below, and the third fraction (MW: ≤ 5 kDa) having the best ACE inhibitory activity was subjected to gel filtration. Gel filtration was performed by dissolving the third fraction (MW: ≤ 5 kDa) at 100 mg/mL using DW in a Sephadex G-10 column (φ2.5 X 100 cm) and injecting 3 mL. Distilled water was eluted by flowing at a flow rate of 2 mL/min, and absorbance was measured at 220 nm using OPTIZEN ^TM POP. According to the measured absorbance, the eluted four fractions (F1 to F4) were recovered (FIG. 2), and the ACE inhibitory activity was evaluated for these four fractions as in the experimental example below, and the third having the most excellent ACE inhibitory activity. For fraction F3, using RP-HPLC equipped with an ODS C18 column (Waters, Atlantis T3 column, 3.0 Х 150 mm, 3 μm), and a linear phase gradient method ((0 min-30 min) with acetonitrile and distilled water solution : 0:100 v/v-40:60 v/v, 30-40 min: 50:50 v/v, 40-50 min: 100:0 v/v and 50-60 min: 100:0 v/v ) Was eluted at a flow rate of 0.3 mL/min, and the absorbance was measured at 220 nm and 280 nm and monitored. As a result, a peark was clearly identified, and a total of three single peptides were fractionated as shown in Table 1 below.

For three single peptides, amino acid sequences were analyzed with a Q-TOF ESI mass spectrometer (Bruker Daltonics, 255748 Germany) and are shown in Table 1 below, and each analysis result is shown in FIGS. 3 to 5. In addition, as a result of comparing the molecular weight measured with a Q-TOF ESI mass spectrometer (Bruker Daltonics, 255748 Germany) (FIGS. 6 to 8) and the molecular weight of the theoretical value, it was confirmed that all three peptides were identical.

<Experimental Example> Evaluation of antihypertensive activity

1. Experimental method

1.1 Evaluation of ACE inhibitory activity

The evaluation of ACE inhibitory activity was performed in 3-hydroxybutyryl-Gly-Gly-Gly, a substrate, according to the method presented in Kang et al. (Kang N, et al., ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY, 2015, 39, 764-772). The enzyme activity was measured by determining the amount of hyroxybutylic acid.

Specifically, ACE kit-WST (Dojindo Inc., Japan) was used, and 50 μL of ACE coenzyme solution was added to 50 μL of the sample solution according to the manufacturer's protocol, followed by pre-reaction at 37°C for 5 minutes, and 50 μL of the substrate was added. It was reacted again at 37°C for 1 hour. Then, 150 μL of 1 N HCl was added to stop the reaction, 750 μL of ethyl acetate was added, stirred for 1 minute, centrifuged (4oC, 5,000 rpm, 10 min), and 500 μL of supernatant was obtained. The supernatant was completely dried at 120oC for 30 minutes, 2 mL of methanol was added, and the absorbance was measured at 228 nm. As a control, 50 μL of the extraction solvent was added instead of the extract, and the inhibitory activity was calculated using the formula, ACE inhibition (%) = [1-{(CAbs-SAbs)/(CAbs-BAbs)}]×100: "CAbs: control absorbance, SAbs: sample absorbance, BAbs: absorbance without sample added"

1.2 Statistical Analysis

Statistical treatment of the experimental results was expressed as the mean ± standard deviation for each sample. One-way ANOVA-test was conducted using the SPSS program (SPSS Inc. Ver. 12.0), and the significance of the survey items was verified by Turkeys multiple range test. P-value compared to the control group *; p<0.05, **; It was expressed as p<0.005.

2. Experiment result

Regarding the evaluation results of the ACE inhibitory activity, the ACE inhibitory activity (IC ₅₀ , concentration corresponding to 50% inhibitory activity) of the protamex hydrolyzate of the Big Valley hippocampus is shown in Table 2 below along with the yield of the hydrolyzate. Table 3 for the ACE inhibitory activity of the fractions according to the three molecular weights of the hydrolyzate, and for the four gel filtration fractions (F1 to F4) of the third fraction (MW: ≤ 5 kDa) among the fractions, in Table 4, gel filtration The three peptides isolated from the third fraction, F3 among the fractions, are shown in Table 5, respectively.

Referring to Table 2, the ACE inhibitory activity of the protamex hydrolyzate of the Big Valley hippocampus was found to have an IC ₅₀ of 0.014 mg/ml, and referring to Table 3, the third fraction (MW: ≤ 5 kDa) of IC ₅₀ was 0.044 mg/ml, showing the best ACE inhibitory activity. Referring to Table 4, among the four gel filtration fractions (F1 to F4) of the third fraction (MW: ≤ 5 kDa), the third fraction, F3, had an IC ₅₀ of 0.330 mg/ml, showing the most excellent activity, 3 Among the canine peptides, the second peptide was the most active.

<110> HAECHUNMA <120> Composition for Anti-hypertension Using Protamex Hydrolysates of Hippocampus abdominalis, Fractions Thereof, or Effective Peptides Thereof <130> PP19-665 <160> 3 <170> KoPatentIn 3.0 <210> 1 <211> 4 <212> PRT <213> Unknown <220> <223> Hippocampus abdominalis <400> 1 Ala Pro Thr Leu One <210> 2 <211> 7 <212> PRT <213> Unknown <220> <223> Hippocampus abdominalis <400> 2 Cys Asn Val Pro Leu Ser Pro 1 5 <210> 3 <211> 5 <212> PRT <213> Unknown <220> <223> Hippocampus abdominalis <400> 3 Pro Trp Thr Pro Leu 1 5

Claims (6)

A composition for improving hypertension comprising a fraction of the Big Valley hippocampus protamex hydrolyzate, or any one of the three peptides in Table 1 below, or a pharmaceutically acceptable salt of the peptide
[Table 1]

The method of claim 1,
The fraction is a composition, characterized in that the fraction having a molecular weight of 5 kDa or less when the supernatant of the Big Valley hippocampus protamex hydrolyzate is fractionated according to molecular weight.
The method of claim 1,
The composition, characterized in that the peptide is the second peptide of Table 1.
The method according to any one of claims 1 to 3,
The composition is a composition, characterized in that the food composition.
The method according to any one of claims 1 to 3,
The composition is a composition, characterized in that the pharmaceutical composition.
The peptides of Table 1 below or a pharmaceutically acceptable salt thereof.
[Table 1]

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