Pharmaceutical compositions for improving blood pressure comprising violet extract

Abstract

The present invention relates to a pharmaceutical composition for the improvement, prevention, or treatment of blood pressure comprising violet extract as an active ingredient. More specifically, the present invention relates to a pharmaceutical composition comprising violet extract as an active ingredient, which is highly useful for the improvement, prevention, and treatment of blood pressure due to its angiotensin-converting enzyme (ACE) inhibitory activity.

Description

Technology Field

[0001] The present invention relates to a pharmaceutical composition for the improvement, prevention, or treatment of blood pressure comprising violet extract as an active ingredient. More specifically, the present invention relates to a pharmaceutical composition comprising violet extract as an active ingredient, which is highly useful for the improvement, prevention, and treatment of blood pressure due to its angiotensin-converting enzyme (ACE) inhibitory activity. Background Technology

[0002] Blood circulation refers to the movement of blood through blood vessels to various parts of the body. Blood supplies oxygen and nutrients to the body's tissues, removes waste products produced by cells, transports hormones to their sites of action, and defends cells against harmful external substances. Additionally, it plays a role in maintaining homeostasis within the body, such as by maintaining an appropriate body temperature and stopping bleeding.

[0003] Endothelial cells of blood vessels play a role in regulating blood fluidity, flow, vascular tone, and arterial function by synthesizing and secreting various types of substances, such as substances that break down blood clots, or by participating in platelet aggregation, thrombus formation, and dissolution.

[0004] When endothelial cells are damaged by inflammatory stimuli, the secretion of water-soluble adhesion molecules increases excessively. Oxidized cholesterol accumulates at the damaged site to form plaque, causing the inner wall of the blood vessel to swell and eventually narrow, obstructing blood flow. When blood vessels are damaged, a hemostatic response begins; however, if the adhesiveness and aggregation of platelets become excessive during this process, the diameter of the blood vessel narrows, potentially leading to circulatory abnormalities. Furthermore, the promotion of blood clot formation within the vessel can result in circulatory problems.

[0005] It is known that impaired blood flow increases the risk of developing cerebrovascular and cardiovascular diseases such as arteriosclerosis and stroke. Diseases of the circulatory system are one of the top three causes of death in South Korea, showing an increasing trend since 2008, and as of 2018, they accounted for approximately 10.7% of the total mortality rate, ranking second.

[0006] Violet (Sweet Pine Scent) Viola canescens The plant has an external shape that is curved like a bow; the outer surface is dark brown, and the cut surface is yellowish-brown. The outer surface is wrinkled, the taproot is long and thick with fibrous roots attached, and the texture is soft and easily broken. The cortex is dark brown, split into segments, and the wood is yellowish-white. The outer surface is yellowish-brown to brown, glossy, and hard, with 5 to 6 longitudinal gills and irregular gills in between. Originating from the Himalayas, the violet has been known since ancient times as an ingredient for painkillers and is also known as nard, one of the perfumes used in Jewish weddings.

[0007] In Korean traditional medicine, these violets are known to have effects that relieve a feeling of fullness, abdominal pain, and lower energy; they are also used to treat age spots, freckles, boils accompanied by nutritional deficiencies, tooth decay, and hemorrhoids; they help restore vitality and clear blockages of qi; and they are also used externally for beriberi and knee edema. Furthermore, violet extracts are known to be used for the prevention and treatment of acute pancreatitis (Korean Publication 10-2010-0095838), and it has been reported that they prevent and treat type 1 diabetes by protecting pancreatic β-cells through the suppression of cytokine-induced immune responses (Korean Publication 10-2011-0098500).

[0008] The inventors of the present invention are violets ( Viola canescens The present invention was completed by newly confirming that the extract exhibits angiotensin-converting enzyme (ACE) inhibitory activity and is effective in improving, preventing, and treating blood pressure. The problem to be solved

[0009] The object of the present invention is to provide a pharmaceutical composition for the improvement, prevention, and treatment of blood pressure.

[0010] Another objective of the present invention is to provide a method for improving, preventing, and treating blood pressure. means of solving the problem

[0011] As one embodiment of the present invention for achieving the above objective, a pharmaceutical composition for improving, preventing, or treating blood pressure is provided, comprising violet extract as an active ingredient.

[0012] The present invention will be described in detail below.

[0013] In one embodiment of the present invention, a pharmaceutical composition for improving blood pressure is provided, comprising violet extract as an active ingredient.

[0014] In one embodiment of the present invention, the extract described above is an extract of a C1-C4 alcohol, preferably a 70% ethanol extract.

[0015] In this invention, the violet may be purchased from a commercial market, or may be collected from nature or cultivated. In this invention, the entire plant of the violet, including leaves, stems, fruits, roots, and the whole plant, may be used.

[0016] In the present invention, the terms “improvement” or “prevention” refer to any act of suppressing the rise in blood pressure or delaying the onset of the disease through the administration of a composition.

[0017] In the present invention, the term "treatment" refers to any act of improving or beneficially altering symptoms caused by blood pressure through the administration of a composition.

[0018] In the present invention, "extract" refers to an active ingredient isolated from a natural product, and specifically refers to a substance isolated from violets. The method for preparing the extract described above may be a conventional extraction method in the art, such as ultrasonic extraction, filtration, and reflux extraction. Preferably, the extract may be obtained by extracting violets, from which impurities have been removed by washing and drying, with water, a C1-C4 alcohol (e.g., methanol, ethanol, propanol, butanol, etc.), or a mixed solvent thereof. More preferably, the extract used in the present invention is an ethanol extract, and most preferably, a 70% ethanol extract. It is preferable that the extraction solvent be 2 to 20 times the dry weight of the violets. After extraction with a solvent, additional methods such as concentration or freeze-drying may be performed.

[0019] In one embodiment of the present invention, the violet extract is characterized by inhibiting angiotensin-converting enzyme (ACE) to prevent the production of angiotensin II. Specifically, the violet extract of the present invention inhibits angiotensin-converting enzyme (ACE) to prevent the production of angiotensin II, and can be used to improve blood pressure or prevent / alleviate or treat symptoms of hypertension.

[0020] In one embodiment of the present invention, the term "hypertension" refers to a condition in which systolic blood pressure is 140 mmHg or higher or diastolic blood pressure is 90 mmHg or higher and persists; it is called the "silent killer" because there are no distinct symptoms in the early stages. However, if left untreated, it is a chronic metabolic disease that causes continuous damage to the blood vessel walls, leading to fatal target organ damage such as stroke, myocardial infarction, and renal failure. Hypertension is caused by various factors including genetic factors, dietary habits, and aging, and in particular, an imbalance in the body's renin-angiotensin system (RAS) is cited as one of the major causes. When ACE activity becomes abnormally high due to aging or obesity, angiotensin II is overproduced, which acts as a direct trigger for the rise in blood pressure.

[0021] In the present invention, the activity of violet extract in improving blood pressure or preventing / alleviating or treating symptoms of hypertension can be confirmed through the angiotensin-converting enzyme (ACE) inhibitory activity of the following examples.

[0022] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. The composition including the pharmaceutically acceptable carrier may be in various oral or parenteral dosage forms. When formulating, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with one or more compounds. In addition, lubricants such as magnesium stearate and talc are also used in addition to simple excipients.

[0023] Liquid preparations for oral administration include suspensions, oral liquids, emulsions, and syrups; in addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspension solvents. Witepsol, Macrogol, Tween 61, cocoa paste, laurin paste, and glycerogelatin may be used as bases for suppositories.

[0024] The above pharmaceutical composition may have any one formulation selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, liquid formulations, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories.

[0025] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.

[0026] In the present invention, the term "pharmaceuticalally effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including individual type and severity, age, gender, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. It may also be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by considering all of the above factors, and this can be easily determined by a person skilled in the art.

[0027] The pharmaceutical composition of the present invention may be used alone or in combination with methods using surgery, hormone therapy, drug therapy, and biological response modulators for the prevention or treatment of blood pressure.

[0028] Meanwhile, the present invention provides a method for improving, preventing, or treating blood pressure, comprising the step of administering a composition for improving, preventing, or treating blood pressure in a pharmaceutically effective amount to an individual who has developed or is likely to develop a blood pressure-related disease.

[0029] In the present invention, the term "individual" refers to any mammal, including humans, that is already in a state of hypertension or is at risk of developing hypertension, and the said disease can be effectively prevented and treated by administering a composition containing an extract or fraction of the present invention to the individual. For example, a human in a state of hypertension can be treated with the pharmaceutical composition of the present invention. Furthermore, the pharmaceutical composition of the present invention can be administered in conjunction with existing antihypertensive agents or therapeutic agents.

[0030] The administration route of the above pharmaceutical composition may be any general route as long as it can reach the target tissue. Depending on the purpose, the pharmaceutical composition of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, orally, intranasally, intrapulmonaryly, or rectally, but is not limited thereto. Additionally, the above pharmaceutical composition may be administered by any device capable of delivering the active substance to target cells.

[0031] The pharmaceutical composition of the present invention is not limited to these, but may be formulated and used in the form of oral formulations such as powders, granules, capsules, tablets, and aqueous suspensions, as well as topical preparations, suppositories, and sterile injectable solutions, according to conventional methods. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. For oral administration, the pharmaceutically acceptable carrier may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavors, etc.; for injectable preparations, it may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc., in combination; and for topical administration, it may include bases, excipients, lubricants, preservatives, etc. The formulations of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injectables, it can be manufactured in the form of unit dosing ampoules or multiple dosing ampoules. In addition, it can be formulated into solutions, suspensions, tablets, capsules, sustained-release formulations, etc.

[0032] Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. Additionally, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. may be additionally included.

[0033] The routes of administration of the pharmaceutical composition according to the present invention include, but are not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal. Oral or parenteral administration is preferred.

[0034] In the present invention, 'parenteral' includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intradural, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0035] The pharmaceutical composition of the present invention may vary depending on several factors including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, elimination rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art, depending on the patient's condition, body weight, degree of disease, drug form, route of administration, and duration, and may be administered at a dose of 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. The administration may be performed once a day or divided into several doses. The dosage does not limit the scope of the present invention in any way.

[0036] In one embodiment of the present invention, a health functional food composition for improving blood pressure or alleviating symptoms of hypertension is provided, containing violet extract as an active ingredient. This composition is characterized by inducing blood pressure regulation by inhibiting the production of angiotensin II through angiotensin-converting enzyme (ACE) inhibitory activity.

[0037] The food composition according to the present invention can be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, chewing gum, ice cream, vitamin complexes, health supplements, etc.

[0038] The food composition of the present invention may include, as an active ingredient, not only the violet extract but also ingredients that are typically added during food manufacturing, such as, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the carbohydrates described above include monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as conventional sugars like dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents, natural flavoring agents [taumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (saccharin, aspartame, etc.) may be used. For example, when the food composition of the present invention is manufactured into a drink or beverage, in addition to the peptide composed of the amino acids of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included.

[0039] The amount of violet extract added to the food composition of the present invention cannot be uniformly specified as it varies depending on the type of food composition to be used, but it may be added within a range that does not impair the original taste of the food, and may typically be in the range of 0.01 to 50 weight%, preferably 0.1 to 20 weight%, relative to the food to be used. In addition, in the case of health functional foods in the form of pills, granules, tablets, or capsules, it may typically be added within the range of 0.1 to 100 weight%, preferably 0.5 to 80 weight%. In one embodiment, the health functional food of the present invention may be in the form of pills, tablets, capsules, or beverages. Effects of the invention

[0040] The pharmaceutical composition containing violet extract according to the present invention as an active ingredient is a composition derived from natural products that has excellent effects in improving, preventing, and treating blood pressure without toxicity or side effects, and can therefore be applied more safely for the improvement and treatment of blood pressure, etc. Brief explanation of the drawing

[0041] Figure 1 is a graph analyzing the angiotensin-converting enzyme (ACE) inhibitory efficacy of a composition containing violet extract of the present invention. Specific details for implementing the invention

[0042] Hereinafter, the specific configuration and operation of the present invention will be explained in more detail through examples and experimental examples. However, the following examples and experimental examples are merely illustrative of the present invention and are not limited thereto.

[0043] Preparation Example 1: Extraction Method

[0044] Dried violets ( Viola canescens)Plant (80 g) was extracted with 70% ethanol at room temperature for 72 hours. The extract was filtered through filter paper and evaporated under reduced pressure at 37°C using a rotary evaporator (Heidolph Hei-VAP Rotary Evaporator). The resulting extract was stored at 4°C until analysis. The extraction yield relative to the weight of the plant part used was calculated as a percentage of the dry weight.

[0045] Other plant extracts tested in comparative experiments with violets were also extracted under the same conditions as above. Dried violets ( Viola canescens) A portion of the plant (10g) was added to 20% ethanol, and the same process was repeated for extraction.

[0046] Experimental Example 1: Preparation of Angiotensin-Converting Enzyme (ACE) and Substrate (HHL) Solutions

[0047] 1 g / 10 ml (w / v) of enzyme source obtained from freeze-dried rabbit lung tissue powder was prepared in 0.1 M sodium borate buffer (pH 8.3) containing 0.3 M NaCl and stored at 4°C for 24 hours. After centrifugation at 15,000 rpm for 60 minutes at 4°C, the supernatant was taken as the crude enzyme solution and stored at -20°C until use. The substrate, hypuryl-histidyl-leucine (HHL), was dissolved in borate buffer (0.5 mM) and stored until use.

[0048] Experimental Example 2: In Vitro Analysis of Angiotensin-Converting Enzyme (ACE)

[0049] The analysis was performed by mixing 150 μl of borate buffer (pH 8.3) substrate solution with 40 μl of inhibitor solution, adding the positive control captopril (or 20% DMSO for the control) and 30 μl of ACE solution, and incubating at room temperature for 5 minutes. After 5 minutes of incubation, 150 μl of the substrate hypuryl-histidyl-leucine (HHL) was added, and the mixture was incubated in an oven at 37°C for an additional 30 minutes. After 30 minutes, 100 μl of 1N HCl was added to stop the reaction, followed by the addition of 700 μl of ethyl acetate to the solution. The supernatant was separated by centrifugation for 1 minute, transferred to a new tube, and evaporated at 65°C for 2 hours. After obtaining the dry powder, 1 ml of borate buffer (pH 8.3) was added, and the degree of absorption at a wavelength of λ = 228 nm was analyzed using UV light.

[0050] Experimental Example 3: Analysis of Angiotensin-Converting Enzyme (ACE) Inhibitory Efficacy Using HPLC

[0051] The analysis was performed by mixing 150 μl of borate buffer (pH 8.3) substrate solution with 40 μl of inhibitor solution, adding the positive control captopril (or 20% DMSO for the control) and 30 μl of ACE solution, and incubating at room temperature for 5 minutes. After 5 minutes of incubation, 150 μl of the substrate hypuryl-histidyl-leucine (HHL) was added, and the mixture was incubated in an oven at 37°C for an additional 30 minutes. After 30 minutes, 500 μl of MeOH was added to stop the reaction, and the solution was centrifuged for 1 minute to obtain the clear liquid from the top for subsequent HPLC analysis.

[0052] HPLC analysis was performed using an Agilent chromatography system on a C18 column (250 mm, 4.6 mm ID) with a particle size of 5 μm, and the analyte was detected at a wavelength of λ = 228 nm. The column was eluted using two solvent systems: (A) 0.05% TFA in water and (B) 0.05% TFA in acetonitrile at a flow rate of 0.7 mL / min and an injection volume of 10 μl.

[0053] The gradient consisted of 20% B for 0 to 5 minutes, was maintained at 100% B for 15 minutes, and then returned to 20% B for 25 minutes.

[0054] Experimental results

[0055] As a result of the above experiments, as shown in Fig. 1, it was confirmed that violet extract exhibits inhibitory activity against angiotensin-converting enzyme (ACE), an enzyme associated with blood pressure regulation. Captopril, used as a positive control, showed ACE inhibitory activities of 90.1%, 69.1%, and 41.5% at concentrations of 540, 324, and 108 μg / ml, respectively, and IC50 50 The value was confirmed to be 150.03 μg / ml.

[0056] Meanwhile, violet extract exhibited ACE inhibitory activities of 80.3%, 59.3%, and 13.4% at concentrations of 1081, 540, and 108 μg / ml, respectively, and IC50 50 The value was confirmed to be 384.79 μg / ml. From the above results, it was confirmed that violet extract exhibits ACE inhibitory activity and can be utilized as an active ingredient in a composition for blood pressure regulation.

[0057] Sample Concentration (ug / ml) ACE inhibition (%) IC 50 (μg / ml) captopril 540 90.1 150.03 324 69.1 108 41.5 Violet extract 1081 80.3 384.79 540 59.3 108 13.4

Claims

Claim 1 A pharmaceutical composition for improving blood pressure containing violet extract as an active ingredient. Claim 2 A pharmaceutical composition for improving blood pressure according to claim 1, characterized in that the extract is a C1-C4 alcohol extract. Claim 3 A pharmaceutical composition for improving blood pressure according to paragraph 2, characterized in that the extract is a 70% ethanol extract. Claim 4 A pharmaceutical composition for improving blood pressure according to claim 1, further comprising one or more auxiliary agents selected from the group consisting of a carrier, an excipient, a disintegrant, a sweetener, a leavening agent, a lubricant, a lubricant, a flavoring agent, an antioxidant, a buffer, a diluent, a dispersant, a binder, and a lubricant. Claim 5 A pharmaceutical composition for improving blood pressure, characterized in that, in claim 1 or 4, the pharmaceutical composition is an oral composition. Claim 6 A pharmaceutical preparation comprising the composition of any one of claims 1 to 4. Claim 7 A health functional food composition for improving blood pressure or alleviating symptoms of hypertension, containing violet extract as an active ingredient. Claim 8 A health functional food composition according to claim 7, characterized in that the composition induces blood pressure regulation by inhibiting the production of angiotensin II through angiotensin-converting enzyme (ACE) inhibitory activity. Claim 9 A health functional food composition according to claim 7, characterized by further comprising one or more auxiliary agents selected from the group consisting of proteins, carbohydrates, fats, nutrients, seasonings, and flavorings.

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