WO2013129769A1 - Composition destinée à traiter ou prévenir l'hypertension artérielle - Google Patents

Composition destinée à traiter ou prévenir l'hypertension artérielle Download PDF

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Publication number
WO2013129769A1
WO2013129769A1 PCT/KR2012/011835 KR2012011835W WO2013129769A1 WO 2013129769 A1 WO2013129769 A1 WO 2013129769A1 KR 2012011835 W KR2012011835 W KR 2012011835W WO 2013129769 A1 WO2013129769 A1 WO 2013129769A1
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Prior art keywords
extract
compound
formula
epoxide hydrolase
composition
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PCT/KR2012/011835
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English (en)
Korean (ko)
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마승진
천길용
류동영
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목포대학교 산학협력단
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Priority claimed from KR1020120021519A external-priority patent/KR101819606B1/ko
Priority claimed from KR1020120031444A external-priority patent/KR101400735B1/ko
Application filed by 목포대학교 산학협력단 filed Critical 목포대학교 산학협력단
Publication of WO2013129769A1 publication Critical patent/WO2013129769A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/84—Valerianaceae (Valerian family), e.g. valerian
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives

Definitions

  • the present invention relates to a composition for treating or preventing hypertension.
  • Hypertension is the most frequent disease among chronic circulatory diseases, and it is relatively asymptomatic, but it can cause fatal complications such as stroke, heart failure, and coronary artery disease.
  • essential hypertension corresponds to 80% of hypertension and is a complex product of genetic and environmental factors, including family history of hypertension, race, salt intake, insulin resistance, obesity, and excessive drinking and aging. It is thought.
  • the epoxide hydrolase plays an important role in the metabolism of harmful foreign compound such as hormones, chemotherapy drugs, calcinegen, environmental pollutants, mycotoxins.
  • the mammalian soluble epoxide hydrolase is a homodimer consisting of two domains. Among them, the C-terminal domain catalyzes the epoxide hydrolysis, the N-terminal domain acts as the phosphatase, and the water-soluble epoxide hydrolase is the epoxyeicosatrienoic acids associated with hypertension. It is known to hydrolyze epoxy fatty acid intermediates such as).
  • the epoxide hydrolase is classified into microsomal epoxide hydrolase (mEH) and soluble epoxide hydrolase (sEH) according to its location and substrate selectivity, and is a plant natural product. It has a complementary function in the decomposition of.
  • the soluble epoxide hydrolase is mainly involved in metabolism of lipid oxides such as arachidonic acid and linoleic acid. Hydrolysis of the epoxyeicosatrienoic acid (EET) of arachidonic acid shows that they can regulate endothelial function by soluble epoxide hydrolase (sEH) by controlling their fusion to coronary endothelial phospholipids.
  • lipid oxides such as arachidonic acid and linoleic acid.
  • EET epoxyeicosatrienoic acid
  • SEH soluble epoxide hydrolase
  • SHR spontaneous hypertensive rats
  • male knockout sEH mice showed significantly lower blood pressure than wild-type mice, demonstrating that soluble epoxide hydrolase can regulate blood pressure.
  • the present invention effectively inhibits the water-soluble epoxide hydrolase, has a blood pressure lowering ability, is useful for the treatment, prevention or improvement of hypertension and does not cause side effects including natural products.
  • the antihypertensive action is excellent, and thus, the substance that inhibits the epoxide hydrolase is used for the treatment, improvement or prevention of cardiovascular diseases or as an active ingredient of the composition for the purpose. Since it can be used as an object, to provide an active ingredient of the composition for inhibiting the epoxide hydrolase.
  • the present invention provides a food composition for improving or preventing cardiovascular diseases, including the extract persimmon extract to achieve the above object as an active ingredient.
  • the present invention also provides a pharmaceutical composition for the treatment or prevention of cardiovascular diseases comprising a compound or a pharmaceutically acceptable salt thereof isolated from the extract of Scent.
  • the present invention provides a food composition for improving or preventing cardiovascular diseases comprising a compound or a pharmaceutically acceptable salt thereof isolated from the extract of Scent.
  • the cardiovascular disease is a disease including heart disease and vascular disease, and in a crowd consisting of hypertension, hypertensive heart disease, heart disease, arrhythmia, stroke, thrombosis, angina pectoris, heart failure, myocardial infarction, atherosclerosis and arteriosclerosis It may be any one selected, preferably hypertension or hypertensive heart disease.
  • blood pressure refers to the force that blood exerts on the vessel wall. When reading blood pressure, it is divided into systolic blood pressure (highest blood pressure) and diastolic blood pressure (lowest blood pressure).
  • systolic blood pressure is the pressure exerted on blood vessels as the heart contracts and releases blood
  • diastolic blood pressure is the pressure the blood vessel receives when the heart receives blood as it expands (relaxes).
  • hypertension is a disease in which the systolic blood pressure is 140 mmHg or more or the diastolic blood pressure is 90 mmHg or more in adults 18 years or older, and the cause is not found. Cases occur, and about 95% of all hypertension patients are essential hypertension. Diseases caused by the hypertension include hypertensive heart disease (hypertensive heart disease).
  • Atherosclerosis refers to a disease in which the walls of the arteries become thick and the elasticity of the arteries is reduced.
  • the blood supply to the organs that received the blood from the hardened arteries is reduced, so that additional diseases may develop.
  • An example of a typical disease related to this is coronary artery disease in which atherosclerosis changes in the coronary arteries that supply blood to the muscles of the heart.
  • the inhibitor of the soluble epoxide hydrolase since the inhibitor of the soluble epoxide hydrolase has an anti-inflammatory or antihypertensive effect, the inhibitory effect of the soluble epoxide hydrolase has the effect of treating, preventing or improving cardiovascular diseases such as hypertension. It is expected to be.
  • the antihypertensive action is excellent, and thus, the substance inhibiting the epoxide hydrolase is used for the treatment, improvement or prevention of a cardiovascular disease, or Can be used as an active ingredient.
  • the present invention relates to a pharmaceutical composition for the treatment or prevention of cardiovascular diseases comprising a compound isolated from the extract of sensation or pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a food composition for improving or preventing cardiovascular diseases comprising a compound or a pharmaceutically acceptable salt thereof isolated from the extract of Scent.
  • examples of the food composition of the present invention include food, food additives, beverages or beverage additives.
  • the present invention provides a composition for inhibiting epoxide hydrolase (EH) comprising a compound isolated from the extract of Persimmon Scent as an active ingredient.
  • EH epoxide hydrolase
  • the epoxide hydrolase may be a soluble epoxide hydrolase (sEH).
  • the inhibitory composition refers to a composition capable of inhibiting the activity of the epoxide hydrolase or inhibiting the activity of the enzyme.
  • the compound is a pharmaceutically acceptable or pharmacological agent, including salts, prodrug conjugates, such as esters and amides, metabolites, hydrates and solvates, etc., as well as the entirety of the specified molecule. Meaning an active derivative.
  • composition is intended to include not only products comprising specified components in specified amounts, but also any products which are produced directly or indirectly from combinations of specified components in specified amounts.
  • pharmaceutically acceptable means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not be detrimental to its recipients.
  • the soluble epoxide hydrolase is a hydroxyeicosatrienoic acid (EETs) in the endothelial cells, smooth muscle and other cell types DHET) is an enzyme that converts to dihydroxy derivatives called.
  • the soluble epoxide hydrolase may be, for example, a human soluble epoxide hydrolase.
  • the inventors of the present invention study the anti-hypertensive substance derived from natural substance or the substance for the treatment or prevention of cardiovascular disease, which is much less likely to be related to the chemically synthesized drug and the possibility of safety related problems such as side effects or resistance is easy.
  • the sapson extract has excellent epoxide hydrolase inhibitory activity
  • the saccharin extract is a safety material used as a food.
  • the methanol extract and the hexane fraction of the methanol extract in the extract of the persimmon extract were more excellent in water-soluble epoxide hydrolase inhibitory effect than when the solvent or fraction solvent.
  • the present invention has been completed based on the above confirmed results.
  • the present invention relates to a food composition for improving or preventing cardiovascular diseases, including the extract of Nardostachyschinensis as an active ingredient.
  • the cardiovascular disease may be hypertension or hypertensive heart disease.
  • the extract may be prepared by a conventional extraction method used in connection with the preparation of plant extracts such as solvent extraction, supercritical extraction, subcritical extraction, soxhlet extraction, SDE extraction.
  • the solvent extraction method means a method of extracting one or two or more of the components in a solid or liquid sample using a solvent in order to extract a specific substance from the plant to be extracted.
  • the supercritical extraction is an extraction method using a supercritical fluid, also called a supercritical fluid extraction (SFE).
  • SFE supercritical fluid extraction
  • the supercritical extraction method uses a supercritical fluid that is a fluid in a state exceeding a critical temperature and a boundary pressure.
  • the supercritical fluid has a high density and has a low viscosity and a remarkably higher diffusion ability than a liquid. Compared to the above, there is an advantage of high efficiency, and carbon dioxide is mainly used as the supercritical fluid.
  • the subcritical extraction method is an extraction method using a fluid similar to a supercritical condition, that is, a critical temperature or a pressure lower than the critical pressure.
  • Subcritical Water Extraction (SWE) using a solvent is mainly used.
  • the steam distillation extraction (SDE) extraction method is an extraction method using a simultaneous distillation extraction apparatus, called a steam distillation method, and is the extraction method most commonly used to extract essential oils. It is a method of extracting essential oils from plants by passing hot steam through the extract and cooling the steam again.
  • the soxhlet extraction method is an extraction method using an soxhlet extraction device, which is mainly used for extracting oils and fats.
  • the persimmon extract is preferably prepared by solvent extraction, supercritical extraction, Soxhlet extraction or SDE extraction.
  • the extract of persimmon is at least one extraction solvent selected from the group consisting of water, alcohol having 1 to 4 carbon atoms of 50% to 100%, ethyl acetate, hexane, chloroform, methylene chloride and dichloromethane, preferably methanol It may be an extract extracted using.
  • the persimmon extract is extracted with methanol, that is, at least one fraction solvent selected from the group consisting of water, alcohol having 1 to 4 carbon atoms, ethyl acetate, hexane and dichloromethane in the extract persimmon methanol, preferably hexane It may be fractionated by.
  • methanol that is, at least one fraction solvent selected from the group consisting of water, alcohol having 1 to 4 carbon atoms, ethyl acetate, hexane and dichloromethane in the extract persimmon methanol, preferably hexane It may be fractionated by.
  • the present invention may be a pharmaceutical composition for the treatment or prevention of cardiovascular diseases comprising a compound or a pharmaceutically acceptable salt thereof isolated from the extract of Persimmon.
  • the present invention is a cardiovascular disease treatment comprising any one compound selected from the group consisting of a compound having a structure of formula (1) and a compound having a structure of formula (2) or a pharmaceutically acceptable salt thereof as an active ingredient Or a prophylactic pharmaceutical composition.
  • the cardiovascular disease may be hypertension or hypertensive heart disease.
  • the present invention may be a composition for inhibiting epoxide hydrolase comprising a compound or a pharmaceutically acceptable salt thereof isolated from the persimmon extract as an active ingredient.
  • the present invention is an epoxide hydrolysis comprising any one compound selected from the group consisting of a compound having a structure of Formula 1 and a compound having a structure of Formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient It may be a composition for inhibiting enzyme.
  • the epoxide hydrolase may be a soluble epoxide hydrolase (sEH).
  • the present invention may be the use of the compound isolated from the extract of the Persimmon scent, specifically for the use of epoxide hydrolase inhibition or for the treatment, improvement or prevention of hypertension.
  • the present invention in this aspect is a compound having the structure of Formula 1 or It may be the use of a compound having a structure of Formula 2.
  • the use of the compound having the structure of Formula 1 or the compound having the structure of Formula 2 may be used for the treatment, prevention or improvement of cardiovascular diseases including anti-inflammatory use or anti-hypertensive use.
  • the extract of Gamsong an active ingredient of the food composition for improving or preventing the cardiovascular disease of the present invention, inhibits the decomposition of epoxy fatty acids such as EETs by inhibiting water-soluble epoxide hydrolase. It can be effective in the improvement or prevention of cardiovascular diseases, and the extract is a natural plant-derived substance unlike the existing cardiovascular disease treatment agent of artificial compound, so the side effects are not a problem, the present invention
  • the extract derived from the extract of the present invention may inhibit NO production and inhibit epoxide hydrolase, the prophylactic or therapeutic effect of cardiovascular disease is recognized, and not only has an anti-inflammatory effect, Since it is a substance derived from a safe plant extract, safety can be ensured with respect to side effects and the like, and thus, it can be used for the treatment, improvement or prevention of cardiovascular diseases, especially hypertension.
  • FIG. 1 is a schematic diagram showing a process for preparing the extract and fractions of the persimmon scent in accordance with an embodiment of the present invention.
  • Figure 2 is a graph showing the yield according to the extraction solvent of the extract persimmon flavor according to an embodiment of the present invention, the horizontal axis represents the extraction solvent, the vertical axis represents the extraction yield.
  • FIG. 3 is a graph confirming the cell viability (%) for confirming the cytotoxicity of the extract persimmon according to an embodiment of the present invention
  • the horizontal axis of the graph means the content of the extract ( ⁇ g / mL)
  • the vertical axis of the graph means cell viability
  • G ⁇ means hydrothermal extract
  • GM means methanol extract
  • GE7 means 70% ethanol aqueous extract
  • GE 95% Mean ethanol extract
  • GH means n-hexane extract
  • GEt means ethyl acetate extract.
  • Figure 4 is a graph (% of control) showing the NO generating ability of the extract according to an embodiment of the present invention as an activity compared to the control, the horizontal axis of the graph means the type of extract, the vertical axis of the graph is NO generating ability
  • GE means 95% ethanol aqueous extract
  • GE7 means 70% ethanol aqueous extract
  • GEt means ethyl acetate extract
  • GH means n-hexane extract
  • G ⁇ stands for hot water extract
  • GM stands for methanol extract.
  • FIG. 5 is a graph showing the water-soluble epoxide hydrolase of the extract persimmon extract according to an embodiment of the present invention (IC 50 ), the horizontal axis of the graph means the type of extract, the vertical axis of the graph is a water-soluble epoxide Mean amount (IC 50 , ⁇ g / mL) to inhibit 50% hydrolase, in each graph, GM means methanol extract, GE means 95% ethanol aqueous solution, GE7 70% Mean ethanol extract, GEt means ethyl acetate extract, GH means n-hexane extract, G ⁇ means hot water extract.
  • IC 50 water-soluble epoxide hydrolase of the extract persimmon extract according to an embodiment of the present invention
  • n-hexane fr Means hexane fraction, chloroform fr.
  • chloroform fr Means chloroform fraction, ethyl acetate fr.
  • butanol Fractions, water fr. Means water fraction.
  • Figure 7 is a photograph showing the TLC results of the hexane fraction of the extract of the persimmon flavor according to an embodiment of the present invention
  • 1 of the horizontal axis of the graph means hexane fraction
  • 3 means sub-fraction 2 of the hexane fraction
  • 4 means sub-fraction 3 of the hexane fraction
  • 4 means sub-fraction 5 of the hexane fraction.
  • Figure 8 shows the epoxide hydrolase inhibitory activity of the column fraction of the sub-fraction 2 of the hexane fraction of the extract of the persimmon extract according to an embodiment of the present invention
  • the horizontal axis of the graph is fraction according to the solvent elution amount of the column It means the type of the result
  • the vertical axis of the graph means the amount of use (IC 50 , ⁇ g / mL) for inhibiting 50% of the water-soluble epoxide hydrolase.
  • Figure 9 is a sub-fraction of No. 2 of the hexane fraction of the extract persimmon extract according to an embodiment of the present invention. Sephadex LH-20 Chromatography of the Elution Fraction Result.
  • FIG. 10 is a photograph showing the Sephadex LH-20 chromatography results of the sub-fraction 3 of the hexane fraction of the extract of Persimmon flavor according to one embodiment of the present invention.
  • FIG 11 is a No. 2 fraction of the hexane fraction of the hexane fraction of extract persimmon according to one embodiment of the present invention.
  • Figure 16 shows the GC-MS spectrum of Compound 1 separated from the eluted fraction.
  • FIG. 12 and 13 are photographs showing the NMR spectral output of Compound 2 separated from sub-fraction 3 of the hexane fraction of the extract of Persimmon Fragrance according to one embodiment of the present invention, and FIG. 12 shows the 1 H-NMR spectral output. It is a photograph, Figure 13 is a photograph showing the result of the 13 C-NMR spectrum.
  • NMR spectrum product of Compound 1 separated from the eluted fraction product Figure 14 is a picture showing the 1 H-NMR spectrum results
  • Figure 15 is a picture showing the 13 C-NMR spectrum results
  • Figure 16 is the EI -MS spectral photograph.
  • the present invention relates to a food composition for improving or preventing cardiovascular diseases, including the extract as an active ingredient.
  • the cardiovascular disease may preferably be hypertension.
  • the persimmon extract may be a persimmon methanol extract, the persimmon extract may be a hexane fraction of the persimmon methanol extract.
  • the food composition may be a functional food composition.
  • the sense songhyang (Nardostachyschinensis) bulriwoomyeo is also known as a perennial plant belonging to the mountains to exchange valerianaceae originating in India or Nepal, fragrant spikenard Reply to this because its taste, spikenard, and Mitch, also known as pine fragrance.
  • the persimmon fragrance is cylindrical in shape with a bow shape, and the outer surface is glossy to yellowish brown to brown, and the cut surface is yellowish brown.
  • the outer surface has 5 to 6 longitudinal folds and irregular folds therebetween, which is hard, long and thick with roots, with beard roots and soft and easy to bend. Cortex divided into dark brown to form pieces, and neck is yellowish white.
  • the sweet scent is said to give a refreshing feeling, bitter taste, spicy and sweet, and in the private sector, it is said to be effective for tooth decay and hemorrhoids. It has been used as an analgesic material since ancient times, and Jews are known as nad fragrance, one of the balms used for weddings.
  • the persimmon incense may be above ground or underground, and may be any one selected from the group consisting of leaves, stems, roots, flowers, fruits, and mixtures thereof.
  • Extract of the present invention may be prepared according to a conventional land plant, for example, root, leaf, stem, flower, fruit or a method for preparing an extract to extract a mixture thereof, solvent extraction method, supercritical extraction method, sub It can be prepared by conventional extraction methods used in connection with the preparation of plant extracts, such as critical extraction, soxhlet extraction, SDE extraction.
  • the solvent extraction method means a method of extracting one or two or more of the components in a solid or liquid sample using a solvent in order to extract a specific substance from the plant to be extracted.
  • the solvent extraction may be cold extraction, hot extraction or thermal extraction, and may be performed using a conventional extraction device, an ultrasonic mill extractor or a fractionator.
  • the supercritical extraction is an extraction method using a supercritical fluid, also called a supercritical fluid extraction (SFE).
  • SFE supercritical fluid extraction
  • the supercritical extraction method uses a supercritical fluid that is a fluid in a state exceeding a critical temperature and a boundary pressure.
  • the supercritical fluid has a high density and has a low viscosity and a remarkably higher diffusion ability than a liquid.
  • the subcritical extraction method is an extraction method using a fluid similar to a supercritical condition, that is, a critical temperature or a pressure lower than the critical pressure.
  • Subcritical Water Extraction (SWE) using a solvent is mainly used.
  • the steam distillation extraction (SDE) extraction method is an extraction method using a simultaneous distillation extraction apparatus, called a steam distillation method, and is the extraction method most commonly used to extract essential oils. It is a method of extracting essential oils from plants by passing hot steam through the extract and cooling the steam again.
  • the soxhlet extraction method is an extraction method using an soxhlet extraction device, which is mainly used for extracting oils and fats.
  • the persimmon extract is preferably prepared by solvent extraction, supercritical extraction, Soxhlet extraction or SDE extraction.
  • the solvent extraction method may be performed by extracting with an extraction solvent or by adding a fractional solvent to the crude extract prepared by extraction with an extraction solvent.
  • the extraction solvent may be at least one selected from the group consisting of water and an organic solvent.
  • the organic solvent may be a polar solvent such as alcohol having 1 to 5 carbon atoms such as methanol or ethanol, ethyl acetate or acetone, and a nonpolar solvent of ether, chloroform, benzene, hexane or dichloromethane, or a mixed solvent thereof.
  • the extraction solvent of the present invention may preferably be at least one solvent selected from the group consisting of water and alcohols having 1 to 4 carbon atoms, and more preferably at least one selected from the group consisting of alcohols having 1 to 4 carbon atoms and hexane. More preferably, ethanol or methanol, most preferably methanol.
  • the extract of the present invention may be a fraction further subjected to the fractionation process after adding a fractional solvent to the crude extract extracted with the solvent.
  • the fractional solvent is a solvent selected from the group consisting of ethyl acetate, ether, chloroform, benzene, hexane, methylene chloride, butanol, water and mixed solvents thereof, preferably hexane, chloroform and a mixture thereof. More preferably, it may be n-hexane.
  • the prepared extract or the fraction obtained by performing the fractionation process can be filtered or concentrated or dried to remove the solvent, it can be carried out both filtration, concentration and drying.
  • the filtration may be performed using a filter paper or a reduced pressure filter
  • the concentration may be concentrated under reduced pressure using a reduced pressure concentrator, for example, a rotary evaporator
  • the drying may be performed by, for example, a lyophilization method.
  • the extract may be obtained by the following method.
  • the extract may be prepared by adding methanol to the root of the extract persimmon, filtered and concentrated under reduced pressure, and then lyophilized.
  • the saenghyang fragrance fraction may be prepared by sequentially adding the fraction solvent of hexane, chloroform, ethyl acetate, butanol and water to the saenghyang extract, and then separating each solvent fraction.
  • the extract of Methanol fragrance extract is not only excellent in inhibiting ability of the water-soluble epoxide hydrolase, but also less likely to cause side effects or resistance as a substance derived from natural products, and has excellent safety. It can be used as an active ingredient of a food composition for the improvement or prevention of cardiovascular diseases, including.
  • the present invention relates to a cardiovascular disease improvement or prevention composition
  • a cardiovascular disease improvement or prevention composition comprising the extract as an active ingredient.
  • the cardiovascular disease is a disease including heart disease and vascular disease, any one selected from the group consisting of hypertension, hypertensive heart disease, heart disease, arrhythmia, stroke, thrombosis, angina pectoris, heart failure, myocardial infarction, atherosclerosis and arteriosclerosis And preferably hypertension or hypertensive heart disease.
  • the hypertension is a blood vessel-related disease, and may be arterial hypertension with high arterial blood pressure.
  • the hypertension may cause arteriosclerosis or hypertensive heart disease.
  • the extract of S. extract is able to effectively inhibit the water-soluble epoxide hydrolase, the blood pressure lowering ability of the extract was confirmed, the extract of S. extract is a cardiovascular disease such as hypertension and heart disease, arteriosclerosis or cerebral hemorrhage. It is evaluated to have a therapeutic, prophylactic or ameliorating effect.
  • the cardiovascular disease improvement or prevention composition comprising the extract of Persimmon as an active ingredient of the present invention may include the extract of 0.001 to 99.99% by weight, preferably 0.1 to 99% by weight based on the total weight of the composition. .
  • the cardiovascular disease improvement or prevention composition comprising the extract as an active ingredient has an excellent effect in that there is no problem of side effects in that it is an extract of a natural substance, and there is an improvement or prevention effect of the cardiovascular disease. .
  • composition for improving or preventing cardiovascular diseases comprising the extract of Persimmon Fragrance of the present invention as an active ingredient may be directly applied to humans.
  • the cardiovascular disease improvement or prevention composition comprising the extract as an active ingredient may include the extract as an active ingredient alone, and according to the other formulation, method and purpose of use additional ingredients, that is, pharmaceutical Or further nutritionally acceptable carriers, excipients, diluents or accessory ingredients.
  • the term "food” means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. It is intended to include all dietary supplements, beverages, food additives and beverage additives.
  • Examples of the food of the present invention include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.
  • the food in the present invention includes special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, and the like, and natural seasonings (eg, ramen soup).
  • the food, health functional food, beverages, food additives and beverage additives may be prepared by conventional manufacturing methods.
  • the health functional food is a biological defense rhythm control, disease prevention and the like having a food group or a food composition that has added value to the food by using physical, biochemical, biotechnological techniques, etc. It means a food that is designed and processed to fully express the gymnastics function on recovery.
  • the dietary supplement may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of dietary supplements.
  • the drink refers to a generic term for drinking to quench thirst or to enjoy the taste and is intended to include a functional beverage.
  • the beverage is not particularly limited to other ingredients other than including the sweet persimmon extract as an active ingredient in the indicated ratio as an active ingredient, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. have.
  • natural carbohydrates examples include monosaccharides such as glucose, fructose and other disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like, and Sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .
  • the ratio of the natural carbohydrate may be generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the food composition of the present invention.
  • the composition of the present invention may further contain a pulp for the production of natural fruit juices, fruit juice drinks, vegetable drinks.
  • the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • These components can be used independently or in combination.
  • the proportion of such additives is not so critical, but may be selected in the range of 0 to 2,000 parts by weight per 100 parts by weight of the persimmon extract of the present invention.
  • the health functional beverage is a biological defense rhythm control or disease prevention of a beverage group or beverage composition which has added value to the beverage by using physical, biochemical and biotechnological techniques to act and express the function of the beverage for a specific purpose.
  • the health functional beverage is not particularly limited to other ingredients except for containing the persimmon extract of the present invention as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. .
  • natural carbohydrates examples include monosaccharides such as glucose, fructose and other disaccharides such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like, and xylitol Sugar alcohols such as sorbitol and erythritol.
  • natural flavoring agents tacumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the proportion of natural carbohydrates is generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention.
  • the amount of the extract persimmon may be included in 0.01 to 15% by weight of the total food weight, the beverage composition is 100 ml It may be included in a ratio of 0.02 to 5 g, preferably 0.3 to 1 g as a reference.
  • Persimmon extract of the present invention inhibits the water-soluble epoxide hydrolase, it is considered that it can be useful in patients with cardiovascular diseases, specifically hypertension or hypertensive heart disease or when there is such a risk.
  • the present invention provides a pharmaceutical composition for treating or preventing cardiovascular diseases comprising a compound or a pharmaceutically acceptable salt thereof isolated from the extract of Persimmon Scent.
  • the cardiovascular disease may be hypertension or hypertensive heart disease.
  • the present invention provides a food composition for improving or preventing cardiovascular diseases comprising a compound or a pharmaceutically acceptable salt thereof isolated from the extract of Scent.
  • the cardiovascular disease may be hypertension or hypertensive heart disease.
  • the food composition may be a health functional food composition.
  • the substance that inhibits the epoxide hydrolase is used for the treatment, improvement or prevention of cardiovascular diseases, or the composition for the purpose. It can be used as an active ingredient of.
  • the present invention may be the use of the compound isolated from the extract of the persimmon flavor, specifically for the use of epoxide hydrolase inhibition or for the treatment, improvement or prevention of hypertension.
  • the use of the compound isolated from the extract of the sweet persimmon may be for the treatment, prevention or improvement of cardiovascular diseases, including anti-inflammatory use or antihypertensive use.
  • the present invention is a cardiovascular disease treatment comprising any one compound selected from the group consisting of a compound having a structure of formula (1) and a compound having a structure of formula (2) or a pharmaceutically acceptable salt thereof as an active ingredient Or to preventive pharmaceutical compositions.
  • the cardiovascular disease may be any one selected from the group consisting of hypertension, hypertensive heart disease, heart disease, stroke, thrombosis, angina pectoris, heart failure, myocardial infarction, atherosclerosis and arteriosclerosis, preferably hypertension or hypertensive heart disease More preferably hypertension.
  • the hypertension is a blood vessel-related disease, and means arterial hypertension with high arterial pressure.
  • the hypertension may cause arteriosclerosis or hypertensive heart disease.
  • the atherosclerosis refers to a disease in which the walls of the arteries are thickened and the elasticity of the arteries is reduced. When the atherosclerosis develops, the blood supply to the organs that received the blood from the hardened arteries is reduced, so that additional diseases may develop.
  • An example of a typical disease related to this is coronary artery disease in which atherosclerosis changes in the coronary arteries that supply blood to the muscles of the heart.
  • a pharmaceutical composition for treating or preventing cardiovascular diseases comprising any one compound selected from the group consisting of the compound having the structure of Formula 1 and the compound having the structure of Formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient It may include one or more of the pharmaceutical diluents selected from saline, buffered saline, dextrose, water, glycerol and ethanol, diluents are not limited thereto.
  • the pharmaceutical composition for treating or preventing the cardiovascular disease may be applied differently depending on the purpose of administration and the disease.
  • the amount of active ingredient administered substantially depends on a variety of related factors, such as the disease to be treated, the extent of the patient's condition, whether it is co-administered with other drugs (eg chemotactic drugs), the patient's age, gender, weight, food, time of administration, It should be determined in consideration of the route of administration and the ratio of the composition.
  • the pharmaceutical composition for treating or preventing the cardiovascular disease may be adjusted according to the dosage form and the severity of the disease, and may be administered once or once or three times a day, but is not limited thereto.
  • the pharmaceutical composition for treating or preventing the cardiovascular disease of the present invention may be administered orally or parenterally.
  • Parenteral administration refers to administration of medications other than oral, namely rectal, intravenous, peritoneal and muscular, arterial, transdermal, nasal, inhaled, ocular, and subcutaneous.
  • the pharmaceutical composition for treating or preventing cardiovascular diseases may be formulated in any form such as oral dosage form, injectable solution or topical preparation.
  • Formulations are preferably prepared for oral and injectable administration (true solution, suspension or emulsion), most preferably in oral form such as tablets, capsules, soft capsules, aqueous pharmaceuticals, pills, granules and the like. desirable.
  • the peptide of the present invention may be filled into a soft capsule without an excipient, and may be made into a suitable formulation after being mixed or diluted with the carrier.
  • suitable carriers include starch, water, saline, Ringer's solution, dextrose and the like.
  • the pharmaceutical composition for treating or preventing the cardiovascular disease may be a pharmaceutical composition.
  • the pharmaceutical composition may be directly applied to animals including humans.
  • the animal is a biological group corresponding to a plant, and mainly ingests organic substances as nutrients, and means that digestion, excretion, and respiratory organs are differentiated, and preferably, vertebrates, more preferably mammals.
  • the mammal may be human, pig, cow or goat, etc., preferably human.
  • the pharmaceutical composition may include any one compound selected from the group consisting of the compound having the structure of Formula 1 and the compound having the structure of Formula 2 or a pharmaceutically acceptable salt thereof as an active ingredient, and Depending on the formulation, the method of use and the purpose of use, it may further comprise additional ingredients, ie, pharmaceutically acceptable or nutritionally acceptable carriers, excipients, diluents or accessory ingredients.
  • the pharmaceutical composition may be added to the nutrients, vitamins, electrolytes, flavors, coloring agents, neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH regulators, and stabilizers in addition to the active ingredient. It may further contain an agent, a preservative, glycerin, alcohol, a carbonation agent used in the carbonated beverage, and the like.
  • the carrier, excipient or diluent may be lactose, dextrose, sucrose, sorbitol, mannitol, ziitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose Selected from the group consisting of loose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, saline It may be one or more, but is not limited to any conventional carrier, excipient or diluent can be used.
  • the components may be added independently or in combination to any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 or a pharmaceutically acceptable salt thereof as the active ingredient. Can be.
  • the pharmaceutical composition is 0.001% to 99.9% by weight of any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 or a pharmaceutically acceptable salt thereof based on the total weight of the composition %, Preferably 0.1% to 99% by weight, more preferably 1% to 50% by weight.
  • the pharmaceutical composition may further include conventional fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers or preservatives when formulated, and oral or parenteral.
  • conventional fillers extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers or preservatives when formulated, and oral or parenteral.
  • solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, water, or the like in the active ingredient. It may be prepared by mixing cross or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.
  • Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups.In addition to the commonly used simple diluents, water and liquid paraffin, various excipients may include, for example, wetting agents, sweeteners, fragrances, and preservatives. have.
  • the formulation of the pharmaceutical composition of the present invention may be in a preferred form depending on the method of use, in particular by adopting methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be formulated. Examples of specific formulations include warnings, granules, lotions, linings, limonades, powders, syrups, ointments, liquids, aerosols, EXTRACTS, elixirs, ointments, liquid extracts, emulsions, Suspensions, premises, acupuncture, eye drops, tablets, suppositories, injections, spirits, capsules, creams, pills, soft or hard gelatin capsules.
  • compositions of the present invention are preferably formulated using appropriate methods known in the art or using methods disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA. Can be converted.
  • the dosage of the pharmaceutical composition according to the present invention may be appropriately selected by those skilled in the art in consideration of the method of administration, the age, sex and weight of the recipient, the severity of the disease, and the like.
  • the pharmaceutical composition of the present invention is 0.000001 mg / kg / day to 1000 mg based on any one compound selected from the group consisting of a compound having a structure of Formula 1 and a compound having a structure of Formula 2 It can be administered as / kg / day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
  • the pharmaceutical composition of the present invention in addition to any one compound selected from the group consisting of a compound having a structure of the formula (1) and a compound having the structure of the formula (2) or a pharmaceutically acceptable salt thereof is a known cardiovascular system
  • the compound may further include a compound having a therapeutic effect on the disease, and may be included in an amount of 5 parts by weight to 100 parts by weight based on 100 parts by weight of the active ingredient.
  • the present invention also provides a food composition for improving or preventing cardiovascular diseases comprising any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 or a salt thereof as an active ingredient. do.
  • a food composition for improving or preventing cardiovascular diseases comprising any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 or a salt thereof as an active ingredient. do.
  • Examples of the food composition of the present invention include food, food additives, beverages or beverage additives.
  • a food composition comprising any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 or a salt thereof as an active ingredient is an appropriate carrier, excipient, and the like commonly used in the preparation thereof. Diluents may be further included.
  • the term "food” means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. It is intended to include all foods, food additives, health functional foods and beverages, preferably gum or candy.
  • any compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 or a salt thereof of the present invention may be added, for example, various foods, drinks, gum , Candy, tea, vitamin complexes, and functional foods.
  • the food in the present invention includes special nutritional products (e.g., crude oil, young, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasonings ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g.
  • fermented milk, cheese, etc. other processed foods
  • kimchi, pickles various kimchi, pickles, etc.
  • beverages examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, ice cream, etc., natural seasonings (eg, ramen soup, etc.), vitamin complexes, alcoholic beverages, alcoholic beverages, and other dietary supplements.
  • the food, beverage or food additives may be prepared by a conventional manufacturing method.
  • Functional food in the present invention is the control of biological defense rhythm, disease prevention and recovery of food groups or food compositions that have added value to the food by using physical, biochemical, biotechnological techniques, etc.
  • the functional food may include food acceptable food additives, and may further include appropriate carriers, excipients and diluents commonly used in the manufacture of functional foods.
  • the present invention provides a composition for inhibiting epoxide hydrolase (EH) comprising a compound isolated from the extract of Persimmon Scent as an active ingredient.
  • EH epoxide hydrolase
  • the epoxide hydrolase may be a soluble epoxide hydrolase (sEH).
  • the inhibitory composition refers to a composition capable of inhibiting the activity of the epoxide hydrolase or inhibiting the activity of the enzyme.
  • the present invention is an epoxide hydrolase comprising any one or a pharmaceutically acceptable salt thereof selected from the group consisting of a compound having a structure of Formula 1 and a compound having the structure of Formula 2 as an active ingredient It may be a composition for inhibiting (epoxide hydrolase, EH).
  • the epoxide hydrolase may be a soluble epoxide hydrolase (sEH).
  • the present invention provides a compound having the structure of Formula 1 or structure of Formula 2 It may be the use of the compound having a. The use may be for the inhibition of epoxide hydrolase.
  • the compound having the structure of Formula 1 or the compound having the structure of Formula 2 may be used for the prevention or treatment of cardiovascular diseases including anti-inflammatory use or anti-hypertensive use.
  • Any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 may be used alone in the composition for inhibiting epoxide hydrolase, and other pharmacologically acceptable Carriers, excipients, diluents or accessory ingredients may further be included.
  • composition comprising any one compound selected from the group consisting of a compound having a structure of Formula 1 and a compound having the structure of Formula 2 is used as a medicament, or used in medicine or pharmaceutical use
  • Any one compound selected from the group consisting of a compound having the structure of Formula 1 and a compound having the structure of Formula 2 may be mixed with a pharmaceutically acceptable carrier or excipient or diluted with a diluent according to a conventional method. Can be.
  • the content of any one compound selected from the group consisting of the compound having the structure of Formula 1 and the compound of Formula 2 in the composition is 0.001% to 99.9%, 0.1% to 99% by weight Or it may be 1% to 20% by weight, but is not limited thereto, the content of the compound may be used to appropriately adjust to the desired content according to the mode of use and method of use of the composition.
  • Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , Dextrin, calcium carbonate, propylene glycol, liquid paraffin, and one or more selected from the group consisting of physiological saline, but is not limited thereto, and any conventional carrier, excipient or diluent may be used.
  • compositions of the present invention can be formulated using any suitable method known in the art or using methods disclosed in Remington's Pharmaceutical Science (Recent Edition, Mack Publishing Company, Easton PA).
  • the dosage of the composition may be determined in consideration of the method of administration, age, sex, severity, condition of the patient, absorption of the active ingredient in the body, inactivation rate and the drug used in combination, for example, based on the daily effective ingredient 0.1 mg / kg (body weight) to 500 mg / kg (body weight), 0.1 mg / kg (body weight) to 400 mg / kg body weight or 1 mg / kg body weight to 300 mg / kg body weight It can be administered, and can be administered once or divided into several times.
  • the dosage does not limit the scope of the invention in any aspect.
  • Persimmon Scent Nardostachyschinensis
  • the root dried dried persimmon scent was purchased as a medicinal herb from Power Ginseng Dang Pharmaceutical Co., Ltd., Sangdong, Mokpo-si, Jeonnam, Korea.
  • the prepared extract was Whatman NO. Filtration under reduced pressure with 2 (Whatman, England) yielded each of filtrate and residue.
  • the obtained residue was repeated twice in the same manner as described above to perform the extraction and filtration process.
  • the hot water extract in the obtained filtrate was freeze-dried (Freezing dryer, Samwon, Korea), and finally a saccharin hot water extract was obtained, and the remaining organic solvent extract was equipped with a cooling aspirator (CA-11, Eyela, Tokyo, Japan). Extract was prepared by vacuum concentration at 35 ⁇ 2 °C using a vacuum evaporator (N-2N, Eyela, Tokyo, Japan).
  • the extraction yield of the extract was measured and shown in FIG. 2.
  • the extraction yield was determined by the weight ratio (w / w,%) of the extract obtained relative to the sweet scent administered in the preparation of the extract.
  • the extraction yield for each solvent of the persimmon aroma was 14.1% for methanol extract (GM), 12.3% for the 95% ethanol aqueous solution extract (GE), and 70% for the extract. 11.7% of the ethanol aqueous solution extract (GE7), 7.1% of the ethyl acetate extract (GEt), 7.2% of the n-hexane extract (GH).
  • the yield of the hydrothermal extract was 8.7%.
  • a Nardostachyschinensis fraction In order to prepare a Nardostachyschinensis fraction, first, a methanol extract was prepared.
  • Example 1-1 20 L of methanol was added to 3.0 kg of the persimmon incense roots in the cold storage of Example 1-1, and ground with a homogenizer (BM-2 Nissei bio-mixer, Nihonseiki Kaiseiki LTD, Japan), as in Example 1-1, 3 Extraction was repeated several times to obtain a total of 60 L of extract of persimmon root methanol.
  • BM-2 Nissei bio-mixer Nihonseiki Kaiseiki LTD, Japan
  • the methanol extract obtained above was Whatman NO. After filtering under reduced pressure to 2, the filtrate was prepared by extracting the filtrate under a reduced pressure at 35 ⁇ 2 ° C. using a vacuum evaporator equipped with a cooling aspirator to obtain 425 g of extract. The extract was stored at -20 ° C.
  • the solvent fraction of the extract was first dissolved in distilled water 20.0 g (141.2 g dry wt. Eq.) Of methanol extract, then n-hexane (5.9 g), chloroform (10.3 g), ethyl acetate (0.3 g), butanol (1.7 g) and water (0.8 g) were sequentially added to the fractional solvent, followed by solvent fractionation.
  • the obtained fraction was concentrated under reduced pressure in the case of a fraction layer using an organic solvent to obtain a fraction, and the water fraction was lyophilized to obtain a fraction.
  • Fraction yield of the fraction was determined by the weight ratio (w / w,%) of the fraction obtained relative to the sweet smell administered in the extraction step.
  • the fraction of persimmon was obtained 5.9 g for n-hexane, 10.3 g for chloroform, 0.3 g for ethyl acetate, and 1.7 g for butanol.
  • nitric oxide (NO) production for each extract solvent of the extract of Persimmon extract prepared in Example 1, a RAW 264.7 macropharge (mouse) cell line as a cell line was purchased and cultured at ATCC.
  • the RAW 264.7 macropharge (mouse) cell line contains 10% FBS (GIBCO, 16000-044) and 1% penicillin-streptomycin (GIBCO, 15140-122) in an incubator at 5% CO 2 , 95% humidity and 37 ° C. Cultured using a 75 cm 2 T-flask containing the DMEM medium (GIBCO, 12100-046).
  • the cultured cells were used for the experiment while subcultured every two or three days.
  • the cell number of the RAW 264.7 was measured using a hemocytometer made of a single cell.
  • Cytotoxicity of the extract of the persimmon extract prepared in Example 1 was confirmed by the method of cell viability (%) using the RAW 264.7 cells through the MTT assay.
  • the RAW 264.7 cells were suspended in a monolayer using 0.25% trypsin-EDTA, and precultured by dispensing 10% FBS MEM medium to 1 x 10 5 / 200 ⁇ L per well of a 96well plate. After 24 hours, the sample was replaced with fresh medium, and the final sample concentration was 10 ⁇ g / mL, 20 ⁇ g / mL, 50 ⁇ g / mL and 100 ⁇ g / mL, and the total volume was 200 ⁇ L for 72 hours. Incubated.
  • MTT formazan was formed by adding 20 ⁇ L of 2 mg / mL MTT (2,5-diphenyl-2H-tetrazoliumbromide, Sigma, USA) solution (in PBS) and incubating for 3 to 4 hours.
  • the purple MTT formazan was correctly dissolved in 1 mL of DMSO solvent, and absorbance was measured at 540 nm with an ELISA microplate reader (680, BIO-RAD, JAPAN). The measurement results are shown in FIG. 3.
  • the hydrothermal extract (GW), 95% ethanol aqueous solution extract (GE), and 70% ethanol aqueous solution extract (GE7) of the extracts did not show toxicity to cells up to a concentration of 100 ⁇ g / mL
  • methanol extract (GM) it showed 96.8% cell viability at a concentration of 50 ⁇ g / mL, and it was confirmed that it was safe up to a concentration of 50 ⁇ g / mL
  • n-hexane extract (GH) and ethyl acetate extract (GEt) The case showed low virulence with 83% cell viability at a concentration of 100 ⁇ g / mL.
  • the sample treatment concentration of the extract for each extract solvent was determined to be 50 ⁇ g / mL in the experiment to confirm the NO production inhibitory ability and the inhibitory ability of the water-soluble epoxide hydrolase of the extract according to each extract solvent. It was.
  • the extract showed overall NO production inhibiting ability, and the NO production of the relatively polar solvent extract of the hydrothermal extract (GW) and the 70% ethanol aqueous solution extract (GE7)
  • the inhibitory activity was 80.2% and 78.3%, showing a lower inhibitory activity than the other extracts.
  • the non-polar solvent extracts of n-hexane extract (GH), ethyl acetate extract (GEt), methanol extract (GM) NO production was 61.3%, 62.4% and 56.3%, respectively, indicating a high inhibitory activity.
  • the measuring method is specifically as follows. First, 150 ⁇ L of 25 mM bis / TrisHCl buffer (pH 7.0, containing 0.1 mg / mL of BSA) solution was added to a 96 well plate, and 2 ⁇ L of DMSO in the blank well and the DMSO solution prepared in Example 1 were prepared. Each 2 ⁇ L was dispensed.
  • CMNPC solution which is a substrate
  • a fluorescent microplate reader excitation 340 nm, emission 460 nm
  • the IC 50 value of the methanol extract (GM) among the extracts of Saengsonghyang showed the best inhibitory activity at 4.5 ⁇ g / mL, 95% ethanol aqueous solution extract (GE) and n-hexane extract (GH).
  • the IC 50 value of) was 7.0 ⁇ g / mL or 7.5 ⁇ g / mL, respectively, and the IC 50 value of the hydrothermal extract (GW) was 123.5 ⁇ g / mL, respectively, showing the lowest inhibitory activity.
  • Example 2 In order to measure the inhibitory ability of nitric oxide (NO) generation by fractional solvent of the extract of the saenghyanghyang methanol using the optimum extraction solvent identified in Example 2, it was performed in the same manner as in Example 2-2. The measurement results are shown in FIG. 6.
  • the NO production inhibitory activity was highest in the n-hexane fraction, 49.7%, 38.7% in the chloroform fraction, 21.1% in the ethyl acetate fraction, and 11.6% in the butanol fraction. It was found that the water fraction was 1.9%, which was the weakest inhibitor of NO production.
  • Table 1 shows the results of measuring the inhibitory activity of the water-soluble epoxide hydrolase (sEH) by fractional solvents of the extracts of saenghyang-methanol using methanol as the optimal extraction solvent identified in Example 2.
  • SEH water-soluble epoxide hydrolase
  • the IC 50 value of the n-hexane fraction showed the strongest inhibitory activity as 3.5 ⁇ g / mL, the next is the chloroform fraction IC 50 value of 7.0 ⁇ g / mL, while the ethyl acetate fraction
  • the IC 50 values were 21.7 ⁇ g / Ml and 50.3 ⁇ g / mL in the and butanol fractions, respectively.
  • the extract of saenghyanghyang preferably the extract of saenghyanghyang
  • cardiovascular system including blood pressure lowering ability and antihypertensive activity It is suggested that it has a treatment, prevention, or amelioration effect on a related disease.
  • the hexane fraction since the hexane fraction has the highest inhibitory activity of the soluble epoxide hydrolase among the fractions of the extract of Gamsong-hyang, the cardiovascular system including the blood pressure lowering ability and the antihypertensive activity related to the soluble epoxide hydrolase inhibition activity from the above results. It can be confirmed that the disease has a therapeutic effect.
  • the silica gel adsorption column is made of slurry by mixing silica gel (40-63 ⁇ m 60, SILICYCLE, Canada) equivalent to 200 times of the sample with n-hexane: ethyl acetate 99: 1 (v / v). It was prepared by filling in a column (35 mm ⁇ 700 mm). Subsequently, the sample was dissolved in n-hexane: ethyl acetate 99: 1 (v / v) mixed solvent and filled in a column, and then eluted with 1 L of n-hexane: ethyl acetate 99: 1 (v / v) mixed solvent.
  • the Sephadex LH-20 column is prepared by mixing Sephadex LH-20 (17-0090-01, GE Healthcare, Sweden) with chloroform: methanol 1: 1 (v / v) to make slurry, and after 24 hr swelling (30 mm ⁇ 700 mm) was prepared by filling up to 500 mm.
  • the sample fractions were loaded and eluted with 700 mL with a mixed solvent system of chloroform: methanol 1: 1 (v / v).
  • Each eluted sub-fraction was aliquoted into 25 mL test tubes, 15 mL, concentrated to 0.6 mg / dry weight, and concentrated under reduced pressure to measure hsEH inhibitory activity. The measurement results are shown in FIG. 8.
  • the ratio (Ve / Vt) of the elution volumn to the bed volumn is in the range of 0.60 to 0.68.
  • 14 to No. 16 showed 34.6%, 51.1%, and 80.8% of hsEH inhibitory activity, respectively.
  • No. 14 to No. 16 eluted fractionation layer was confirmed by TLC.
  • 16 elution fractions were identified as a single spot. The 16 eluting fractions were selected as fractions for purification by high performance liquid chromatography (HPLC).
  • the purification by HPLC was specifically carried out in the following manner.
  • the column used for HPL was C18 column (10 ⁇ 250 mm, 5C18-AR-II, Nacalai, JAPAN), and 210 nm detection at a flow rate of 4.72 mL / min (Model 980E, JASCO, Japan). It was performed at a wavelength (MD-2010 Multiwavelength detector, JASCO, Japan).
  • the No. The resulting eluted fraction was separated by ODS-HPLC using 50% acetonitrile as a mobile phase, and the main peak was repeatedly extracted from the elution fraction of t R 25.86 minutes to obtain the active compound 1.
  • sub-fraction 3 was repeatedly fractionated from the elution fraction at t R 12.85 minutes by ODS-HPLC using 50% acetonitrile as a mobile phase to obtain compound 2 as an active substance.
  • Example 4-1 In order to confirm the structures of the two effective compounds identified in Example 4-1, GC-MS analysis, LC-MS analysis, and FT-NMR analysis were performed.
  • GC-MS analysis was analyzed by GC-MS (5975C, Agilent Tech., USA) method. Specifically, the analysis conditions were maintained for 3 minutes at an initial temperature of 60 °C, and then raised to 230 °C by 5 °C by 1 minute, and then maintained at this temperature for 20 minutes. Mass spectrometry was performed by ionizing the components separated from GC at 70 eV. The fragmentation patterns were qualitatively analyzed using the Wiely / NIST library. LC-MS analysis, specifically LC-ESI-MS analysis was performed using LC (Agilent 1100 HPLC) equipped with MS (ESI Ion Trap LC-MS System, Bruker, Germany).
  • 2D-NMR shift correlated spectroscopy (COSY), heteronuclear multiple quantum coherence (HMQC), heteronuclear multiple bond correlation (HMBC) was analyzed.
  • COSY shift correlated spectroscopy
  • HMQC heteronuclear multiple quantum coherence
  • HMBC heteronuclear multiple bond correlation
  • GC-MS analysis for the structural analysis of compound 1 showed 222.2 m / z , 207.2 m / z , 189.2 m / z , 179.2 m / z , 161.1 m at 21.86 min at t R.
  • the 15 signals identified in compound 1 is consistent with the signal of the existing patchouli alcohol, it was confirmed that the compound is patchouli alcohol.

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Abstract

La présente invention concerne une composition destinée à traiter, soulager ou prévenir des maladies cardiovasculaires. Les maladies cardiovasculaires peuvent être de l'hypertension artérielle ou des troubles cardiaques hypertensifs. De plus, la présente invention concerne une composition destinée à inhiber l'époxyde hydrolase. Un extrait de Rhizoma Nardostachysis, qui est l'un des principes actifs de la présente invention, soulage ou prévient considérablement les maladies cardiovasculaires en raison d'excellents effets inhibiteurs de l'époxyde hydrolase soluble et d'effets inhibiteurs notables de génération de NO, et est excellent en ce qu'il n'a aucun effet secondaire dans la mesure où une substance naturelle est contenue comme principe actif. D'autre part, un composé isolé d'un extrait de Rhizoma Nardostachysis parmi les matières actives de la présente invention montre des effets inhibiteurs notables de génération de NO et inhibe l'époxyde hydrolase de manière à pouvoir inhiber l'hydrolyse d'une forme active intermédiaire d'acide gras d'époxy, telle que les acides époxyeicosatriénoïques (EET), qui est associée à l'hypertension et, par conséquent, peut présenter une efficacité dans le traitement et la prévention de maladies cardiovasculaires, y compris un effet antihypertenseur.
PCT/KR2012/011835 2012-02-29 2012-12-31 Composition destinée à traiter ou prévenir l'hypertension artérielle WO2013129769A1 (fr)

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KR10-2012-0021519 2012-02-29
KR1020120021519A KR101819606B1 (ko) 2012-02-29 2012-02-29 감송향 추출물을 함유하는 고혈압 예방 또는 개선용 식품 조성물
KR1020120031444A KR101400735B1 (ko) 2012-03-28 2012-03-28 고혈압 예방 또는 개선용 조성물
KR10-2012-0031444 2012-03-28

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CN108721257A (zh) * 2017-04-25 2018-11-02 成都中医药大学 百秋李醇在制备内皮非依赖的血管舒张剂中的用途
CN108721257B (zh) * 2017-04-25 2021-09-10 成都中医药大学 百秋李醇在制备内皮非依赖的血管舒张剂中的用途

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