KR101851200B1 - Pharmaceutical composition comprising 3-deoxysappanchalcone for preventing or treating vascular disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention and treatment of vascular diseases containing 3-deoxysappanchalcone, wherein the 3-deoxy sulphacalcone has the activity of HO-1 (heme oxygenase-1) And thus can be usefully used for the prevention or treatment of vascular diseases such as arteriosclerosis, hypertension, vascular restenosis, peripheral vascular occlusion, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage and stroke .

Description

[0001] This invention relates to pharmaceutical compositions comprising 3-deoxysulfanyl chalcone for preventing or treating cardiovascular diseases,

The present invention relates to a pharmaceutical composition for preventing or treating vascular diseases containing 3-deoxysappanchalcone as an active ingredient.

The endothelium of blood vessels plays an important role in maintaining vascular homeostasis in vasoreactivity, platelet activation, leukocyte adhesion, proliferation and migration of smooth muscle cells. HO (heme oxygenases) are proteins that promote oxidation of biologically active substances, including heme carbon monoxide (CO), biliverdin, and ferrous iron, In the case of rodents, there are two mutants derived from different genes. HO-2 is known to continuously express in the body, but the expression of HO-1 is influenced by heme, . ≪ / RTI > In addition, the induction of HO-1 expression has been shown to be effective in the treatment of wounds and diseases of the blood vessels (Feng, J. et al., 2011) and to inhibit arteriosclerosis (Roh, 2008) It has been reported that induction of HO-1 expression using natural phytochemical can prevent or treat vascular diseases, angioplasty after restenosis, and the like. In addition, HO-1 is associated with the treatment of cerebrovascular diseases such as cerebral hemorrhage, cerebral infarction and stroke (Alfieri, A. et al., 2011), the treatment of vascular restenosis, vascular hypertrophy and hypertension and inhibition of vascular smooth muscle cell proliferation (Ryter, SW et al., 2009) and is known to have therapeutic effects on various cardiovascular diseases (Paine, A. et al., 2010).

Caesalpinia sappan lignum is a plant native to Asian countries such as Korea, China, and Vietnam. As a medicinal herb, it is dried heartwood (corewood), and its core has been used as a traditional medicinal material such as natural dyestuffs of red color, antimicrobial agents, anti-inflammatory agents, analgesics and analgesics. In addition to this, it is also known to promote blood circulation and remove blood vessels. It is known that the main ingredients of the joan are brazilin, sappanchalcone and brazilein. On the other hand, the present inventors isolated 3-deoxysappanchalcone from the alopecia extract and found that the compound increases the activity of HO-1 (heme oxygenase-1) and inhibits oxidative stress in blood vessels, , Hypertension, coronary artery disease, and the like. The present invention has been completed based on this finding.

It has not yet been disclosed that 3-deoxy sulphacalcone has a therapeutic effect on vascular diseases. It has not yet been disclosed that the 3-deoxy sulphacalcone has the effect of inhibiting angiogenesis in Korean Patent No. 750657, Korean Patent Laid-Open No. 2008-76343 discloses that the extract of Alaska pollack has an effect of treating arteriosclerosis, and Korean Patent No. 1284838 discloses that the fraction of ethyl acetate in the alum is effective for treating various vascular diseases However, the activity of 3-deoxy sulfenate chalcone is not disclosed in the above prior art documents.

Korean Registered Patent No. 750657 (use of extracts of jojoba and the compounds isolated therefrom, registered on August 13, 2007) Korean Patent Publication No. 2008-76343 (Composition for the prevention and treatment of arteriosclerosis containing extract of alum, Aug. 20, 2008) Korean Registered Patent No. 1284838 (Composition for the prevention or treatment of vascular diseases containing extract of jojoba, registered on Mar. 07, 2014)

Roh JH, Serum bilirubin and atherosclerosis., Korean Ddabetes J., 2008, 32, 301-303. Alfieri, A. et al., Targeting the Nrf2-Keap1 antioxidant defense pathway for neurovascular protection in stroke. J Physiol 2011, 589, 4125-4136. Faffe, D.S. et al., Oncostatin M causes VEGF release from human airway smooth muscle: synergy with IL-1 beta., Am. J. Physiol. Lung Cell Mol. Physiol., 2005, 288, 1040-1048. Feng, J. et al., PI3K and ERK / Nrf2 pathways are involved in oleanolic acid-induced heme oxygenase-1 expression in rat vascular smooth muscle cells. J. Cell. Biochem., 2011, 112, 1524-1531. Kim, H.Y. et al., A methylene chloride fraction of Saururus chinensis induces apoptosis through activation of caspase-3 in prostate and breast cancer cells, Phytomedicine, 2011, 18, 567-574. Paine, A. et al., Signaling to heme oxygenase-1 and its anti-inflammatory therapeutic potential Biochemical Pharmacology, 2010, 80, 1895-1903. Vellimana, A.K. et al., Endothelial nitric oxide synthase mediates endogenous protection against subarachnoid hemorrhage-induced cerebral vasospasm. Stroke, 2011, 42, 776-782. Ryter, S.W. et al., Heme Oxygenase-1 / Carbon Monoxide From Metabolism to Molecular Therapy. American Jounal of Respiratory Cell & Molecular Biology, 2009, 41, 251-260. Woo, A. et al., Piceatannol-3'-O-beta-D-glucopyranoside as an active component of rhubarb activates endothelial nitric oxide synthase through inhibition of arginase activity. Exp. Mol. Med., 2010, 42, 524-532.

The present invention provides a pharmaceutical composition for the prevention or treatment of vascular diseases containing 3-deoxysappanchalcone.

The present invention relates to a pharmaceutical composition for preventing or treating vascular diseases containing 3-deoxysappanchalcone of the following formula (1) as an active ingredient.

[Chemical Formula 1]

The vascular disease may be selected from atherosclerosis, hypertension, vascular restenosis, peripheral vascular occlusion, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage, and stroke.

The 3-deoxy sulphacalcone has an effect of increasing the activity of HO-1 (heme oxygenase-1) in vascular endothelial cells.

In another aspect, the present invention provides a health functional food for preventing or improving vascular diseases containing 3-deoxy sulphacalcone as an active ingredient. The health functional foods may be selected from nutritional products including drinks, alcoholic beverages and vitamin complexes including dairy products including soups, meats, sausages, breads, candies, snacks, noodles, ice cream, soups and ionic drinks.

Hereinafter, the present invention will be described in detail.

The 3-deoxy sulphacalcone is a compound of the formula: Caesalpinia sappan lignum) extract may be used, and those synthesized in a laboratory by a conventional method may be used.

The extract can be extracted with water, a C1-C4 lower alcohol or a mixed solvent thereof. The C1-C4 lower alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol . The water, the C1-C4 alcohol, or the mixed solution thereof used in the production of the alum can extract may be used in an amount of 2 to 20 times, preferably 1 to 10 times, based on the weight of the alum. The extraction conditions of the extract can be 1 to 48 hours at 20 to 100 ° C. In addition, as a conventional method in the art, water of the joinery, a C1-C4 alcohol, or a mixed solution thereof extract is dissolved in water or directly after the addition of n-hexane, methylene chloride, acetone, chloroform, ethyl acetate and n-butanol And the solvent may be further fractionated using at least one solvent selected from the group consisting of The production temperature of the extract or its fractions may be 20 to 50 ° C, but is not limited thereto. The extraction time is not particularly limited, but it is preferable to perform extraction within 10 minutes to 1 day. As the extraction apparatus, a conventional extraction apparatus, an ultrasonic pulverization extractor, or a fractionator may be used. The thus-prepared root extract or fraction can be removed by hot air drying, vacuum drying or freeze drying. In addition, the extract or fraction may be purified by using column chromatography. The chromatography can be carried out using silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

The vascular disease may be a disease selected from atherosclerosis, hypertension, vascular restenosis, peripheral vascular occlusion, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage, and stroke, and various other coronary artery diseases, cardiovascular diseases , Peripheral vascular disease, cerebrovascular disease, aortic disease, and the like.

The pharmaceutical composition for preventing or treating vascular diseases containing 3-deoxy sulphacalcone as an active ingredient according to the present invention can be administered in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols Oral formulations, external preparations, suppositories, and sterile injectable solutions. Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, . In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

The dosage of the pharmaceutical composition will depend on the age, sex, body weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 0.1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or several times. The dose is not intended to limit the scope of the invention in any way. The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. In addition, since the pharmaceutical composition of the present invention is a composition derived from a natural product, it has little toxicity and side effects, and thus can be safely used for long-term administration for preventive purposes.

The present invention also provides a health functional food for preventing or ameliorating a vascular disease comprising the above-mentioned 3-deoxy sulfenate chalcone and a pharmaceutically acceptable food additive. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids, and the health functional foods include dairy products including dairy products, meat, sausages, bread, candy, snacks, noodles, It may include nutritional products, including beverages, alcoholic beverages and vitamin complexes, including ionic beverages.

The present invention relates to a pharmaceutical composition for the prevention and treatment of vascular diseases containing 3-deoxysappanchalcone, wherein the 3-deoxy sulphacalcone has the activity of HO-1 (heme oxygenase-1) And thus can be usefully used for the prevention or treatment of vascular diseases such as arteriosclerosis, hypertension, vascular restenosis, peripheral vascular occlusion, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage and stroke .

FIG. 1 is a graph showing that the production of HO-1 (heme oxygenase-1) is increased in HUVEC due to the treatment of 3-deoxy sulphan chalcone.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

EXAMPLES Example 1: Separation, purification and physicochemical properties of 3-deoxy sulfenate chalcone from copper minerals>

3-Deoxysulfan chalcone was separated from the core (heartwood) of dried Caesalpinia sappan lignum. 10 kg of dried core material was refluxed three times for 4 hours with 30 l of methanol, and the resulting liquid phase was dried to obtain 1358 g of methanol extract. The methanol extract was suspended in 5 L of water and then subjected to solvent fractionation in the same volume of hexane, ethyl acetate and butanol sequentially to obtain a hexane fraction, an ethyl acetate fraction and a butanol fraction. 960 g of the ethyl acetate fraction was loaded on a silica gel column (800 x 200 mm), eluted with increasing polarity in a solvent of chloroform: methanol 50: 1 (v: v) 0: 1 (v: v) A total of 14 small fractions were obtained (f.E1 to f.E14) confirming the eluate by thin layer chromatography. The fraction 9 was eluted again with RPC-18 reverse phase chromatography (500 × 50 mm) and methanol: water (1: 2 [v: v] 1: 0 [v: v]) 15 sub-fractions were obtained (f.E9.1 to f.E9.15), also confirming by thin layer chromatography. The fraction f.E9.1 in the ash fraction was purified by high performance liquid chromatography under the conditions of RPC-18 reverse phase chromatography (250 x 20 mm) and methanol: 0.1% triflourocyte (2: 8 [w: w]) To separate 3-deoxy sulphacalcone at 45.9 min.

On the other hand, the physicochemical properties of 3-deoxy sulphacalcone are as follows.

Color: Yellow bed crystals

Molecular formula: C ₁₆ H ₁₄ O ₄

^{1 H NMR (400 MHz, CD} 3 OD): δ 6.52 (1H, d, J = 2.0 Hz, H-3), 6.46 (1H, dd, J = 2.0, 8.4 Hz, H-5), 7.58 (1H , d, J = 8.4 Hz, H-6), 7.41 (1H, d, J = 15.6 Hz, H-8), 7.56 (1H, d, J = 15.6 Hz, H-9), 7.50 (2H, d , J = 8.4 Hz, H- 2 ', 6'), 6.82 (2H, d, J = 8.4 Hz, H-3 ', 5'), 3.89 (1H, s, 2-OCH 3).

^{13 C NMR (100 MHz, CD} 3 OD): δ 128.2 (C-1), 162.6 (C-2), 100.3 (C-3), 164.6 (C-4), 109.0 (C-5), 121.8 (C-6); C-1 '), 131.5 (C-2', 6 '), 117.0 (C-3', 5 '), 133.3 (C- , 161.2 (C-4 ') , 56.2 (2-OCH 3).

Example 2. Identification of the effect of 3-deoxy sulphacalcone on the expression of HO-1.

HUVEC (human umbilical vein endothelial cells) cells were purchased from Cambrex Bio Science (Charles City, IA, USA), EBM-2 basal media and growth supplement (Cambrex Bio Science) Respectively. HUVEC was treated with 1 to 10 μM of 3-deoxy sulphacalcone for 6 hours and then the cells were harvested and resuspended in cell lysis buffer (50 mM Tris-HCl [pH 7.5], 1 1 mM EDTA, 1 mM phenylmethylsulfonyl fluoride, 10 μg / ml pepstatin A, 10 μg / ml aprotinin, 2 mM benzamidine, 50 mM NaF, 5 mM sodium orthovanadate and 150 mM NaCl) to obtain total protein. The total protein was separated by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and transferred to Hybond-P membrane (Amersham Biosciences, Buckinghamshire, UK). The membrane was blocked with 5% skim milk for 1 hour at room temperature and then reacted with the primary antibody of HO-1 for 2 hours. After that, the peroxidase-conjugated secondary antibody was reacted and the amount of HO-1 protein expression was confirmed using a chemiluminescence system (Intron, Seongnam, Korea). 3-deoxy sulphacalcone was dissolved in dimethyl sulfoxide (DMSO), and the final concentration of DMSO was kept at 0.05% in the cell culture medium.

The results of HO-1 protein expression in HUVEC cells treated with 3-deoxy sulphacalcone were quantitatively shown in FIG. 1. As shown in FIG. 1, the concentration and time of 3-deoxy- And the degree of protein expression of HO-1 was increased.

≪ Example 3: Toxicity test &

Example 3-1. Acute toxicity

This experiment was conducted to investigate the toxicity of the 3-deoxysulfan chalcone of the present invention to an animal body in an acute (within 24 hours) when an excessive amount of the 3-deoxysulfan chalcone of the present invention was consumed in a short period and determine the mortality rate. Twenty ICR mouse lines, which are general mice, were assigned to 10 mice in the control group and 10 mice in the experimental group. Nothing was administered to the control group, and the experimental group was orally administered at a concentration of 2.0 g / kg of the 3-deoxy sulphacalcone of the present invention (about 50 times the amount used in general animal experiments). After 24 hours of administration, the respective mortality rates were examined. As a result, the control group and the test group administered with 3-deoxy sulphacalcone of the present invention at a concentration of 2.0 g / kg survived.

Example 3-2. Organ organs toxicity test in experimental group and control group

In order to investigate the effect of C57BL / 6J mice on the organs (tissues) of the animals, blood samples were collected from the experimental group administered with 3-deoxy sulphacalcone of the present invention and the control group animals, Glutamate-pyruvate transferase (GPT) and BUN (Blood Urea Nitrogen) in blood were measured by Select E (Vital Scientific NV, Netherland). As a result, GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to renal toxicity, showed no significant difference compared to the control group. In addition, liver and kidney were cut from each animal and histological observation was carried out with an optical microscope through a conventional tissue section production process. No abnormal abnormalities were observed.

≪ Use examples 1. Pharmaceutical formulations >

1-1. Manufacture of tablets

20 g of 3-deoxy sulphacalcone of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. To this mixture was added a 10% gelatin solution, which was pulverized and passed through a 14-mesh sieve. This was dried, and a mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into tablets.

1-2. Injection preparation

2 g of the 3-deoxy sulfenate chalcone of the present invention was dissolved in distilled water to make 100 ml. This solution was placed in a bottle and sterilized by heating at 20 DEG C for 30 minutes.

<Usage example 2. Food production example>

2-1. Manufacture of cooking seasonings

The content of 3-deoxy sulphacalcone of the present invention was adjusted to 0.2 to 10% by weight to prepare health-promoting cooking sauce.

2-2. Manufacture of flour food products

A food for improving health was prepared by preparing breads, cakes, cookies, crackers and noodles by using the mixture of 3-deoxysulfan chalcone of the present invention as flour at 0.1 to 5.0 wt%.

2-3. Manufacture of soups and gravies

And 0.1 to 1.0% by weight of 3-deoxy sulphacalcone of the present invention was added to the soup and the juice to prepare a health improvement meat product, noodle soup and juice.

2-4. Manufacture of dairy products

Deoxysulfan chalcone of the present invention was added to milk at a concentration of 0.1 to 1.0 wt%, and various dairy products such as butter and ice cream were prepared using the milk.

&Lt; Use Example 3 >

3-1. Vegetable juice manufacturing

0.5 g of the 3-deoxy sulphacalcone of the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

3-2. Manufacture of fruit juice

0.1 g of the 3-deoxy sulfenate chalcone of the present invention was added to 1,000 ml of apple or grape juice to prepare a health promotion fruit juice.

Claims (6)

A cardiovascular system selected from the group consisting of arteriosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage, and stroke, which comprises 3-deoxysappanchalcone of the following formula (1) A pharmaceutical composition for preventing or treating a disease.
[Chemical Formula 1]

delete The method according to claim 1,
Wherein said 3-deoxy sulphacalcone has an effect of increasing the activity of HO-1 (heme oxygenase-1) in vascular endothelial cells. , And a pharmaceutical composition for preventing or treating cardiovascular diseases selected from stroke. A cardiovascular system selected from the group consisting of arteriosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage, and stroke, which comprises 3-deoxysappanchalcone of the following formula (1) Health functional food for prevention or improvement of disease.
[Chemical Formula 1]

delete 5. The method of claim 4,
The health functional food is selected from nutritional products including beverages including alcoholic beverages and vitamin complexes including dairy products including soups, meats, sausages, breads, candies, snacks, noodles, ice cream, soups and ionic drinks A health functional food for preventing or ameliorating a cardiovascular disease selected from arteriosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, cerebral infarction, cerebral hemorrhage, and stroke.

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