KR101336723B1 - A composition comprising extract of Saururus chinensis or compounds isolated therefrom for treating or preventing vascular diseases - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of vascular diseases containing at least one compound of Saururus chinensis extract or saucchinone and 7-hydroxysauchine isolated therefrom As a trichophytium extract of the present invention, or one or more compounds of sacchinone and 7-hydroxysucchin are excellent in inhibiting the activity of arginase in vascular cells, arteriosclerosis, hypertension, angina pectoris, myocardial infarction, It can be usefully used for the prevention and treatment of vascular diseases such as ischemic heart disease, heart failure, complications after angioplasty, cerebral infarction, cerebral hemorrhage, and stroke.

Description

A composition comprising extract of Saururus chinensis or compounds isolated there from for treating or preventing vascular diseases}

The present invention is 300 seconds ( Saururus chinensis ) relates to a composition for the prevention or treatment of vascular diseases containing extracts or compounds isolated therefrom.

More specifically, the present invention inhibits the activity of arginase, promotes the production of nitric oxide (NO) in vascular endothelial cells, thereby relaxing blood vessels, thereby causing atherosclerosis, hypertension, angina, myocardial infarction, ischemic heart disease, and heart failure. The present invention relates to a composition for the prevention or treatment of vascular diseases containing a tritical sect extract or a compound isolated therefrom that can treat complications, cerebral infarction, cerebral hemorrhage, and stroke, etc., which occur after angioplasty.

Endothelial vessels play an important role in maintaining blood vessel homeostasis for vasoreactivity, platelet activity, leukocyte adhesion, smooth muscle cell proliferation and migration.

NO (nitric oxide) produced by the vascular endothelium is known as a major factor of vascular relaxation. The NO is the most potent vascular protective agent that affects vascular response control, platelet activity, lymphocyte fixation, smooth muscle cell proliferation and migration, and relaxes vascular smooth muscle cell to expand blood vessels, and damaged NO signaling system Vascular diseases such as atherosclerosis and hypertension. Nitric oxide synthase (NOS) is an O ₂ and cofactor nicotinamide adenine dinucleotide phospate (NADPH), flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN) , A group of enzymes that synthesize NO from the terminal nitrogen atom of L-arginine in the presence of heme, tetrahydrobiopterin (BH4), and among the known NOSs, n-NOS (neuronal nitric oxide) synthase), e-NOS (endothelial NOS), and i-NOS (inducible NOS). For NOS, NO synthesis, and vasorelaxation, L-NAME (N ^G -nitro-L-argininemethylester), a NOS inhibitor (nitiric oxide synthase inhibitor), inhibits vasorelaxation. oxide) has been shown to be induced by increased synthesis (Xie et al., 2000).

Arginase, on the other hand, inhibits e-NOS (endothelial NOS) from producing L-arganine as a substrate to produce NO. That is, arginase is an enzyme that degrades L-arginine, which is a substrate, and has a competition relationship with e-NOS for the substrate. In addition, arginase not only uses L-arginine as a substrate for e-NOS, but also decomposes L-arginine into ornithine and urea through the urea cycle.

Arginase currently has two isomers. Arginase I expression is expressed in macrophages, hepatocytes, and vascular smooth muscle cells, including lipopolysaccharide (LPS), tumor necrosis factor-α (TNF-α), and interleukin (IL-13). -13), stimulated by altered oxygen tension, balloon dilatation of coronary arteries, etc. (Modolell et al., 1995; Morris et al., 2004).

The expression and activity of vascular endothelial arginase II are oxidized low-density lipoprotein (OXLDL), lipopolysaccharide (LPS), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and 8-bromo- It is induced by various forms of vascular activators such as 8-bromoguanosine 3 ', 5'-cyclic monophosphate (cGMP) and hypoxia (Ryoo et al., 2006; Nelin et al., 2007).

In addition, the inhibition of arginase actively increases NO production and is effective for normal heart function, as well as atherosclerosis, aging, erectile dysfunction and sickle cell disease, which are abnormal blood vessels. It has been reported to be effective in diabetes, hypertension, etc., and the inhibition of arginase increases NO production in red aortic endothelial cells, bovine lung endothelial cells, and coronary arteries of pigs, and HUVEC (human umbilical vein endothelial cells). Has been shown to be associated with the inhibition of angiogenesis.

In addition, the increased activity of arginase II in atherosclerosis-prone mice can be attributed to impaired endothelial NO production, endothelial dysfunction, and vascular endothelial dysfunction. It has been shown to be associated with vascular stiffness and aortic plaque development in the aorta. In contrast, inhibition of vascular endothelial arginase II or deletion of arginase II gene has been reported to be associated with NO production, vascular endothelial and aortic function recovery, and plaque production. Thus, arginase II is a major target for the prevention or treatment of vascular diseases, including atherosclerosis (Ryoo et al., 2008), and the activation of arginase is impaired vascular endothelial dysfunction, hypertension and lung It has also been shown to affect high blood pressure (systematic and pulmonary hypertension).

Three hundred seconds, Saururus chinensis ) is a perennial herbaceous plant belonging to more than three hundred, growing in the marsh near Hyeopjae, Jeju Island, and is called three hundred seconds because of its white leaves, flowers and roots. The height of the three hundred seconds reaches 50-100cm, the root diameter is white, and the mud extends sideways. The leaves are alternate, long egg-shaped oval, with flat edges. Flowers are bisexual and bloom white in June to August. Water inflorescences (穗狀花序) faced with leaves, 10-15㎝ long, with curly hairs, sagging downwards and straightening. The vesicles are egg-shaped round, about 1.5mm in diameter, the pedicel is 2.3mm in length, without petals. Fruits are round and one seed is contained in each thread. In Oriental medicine, dried herbs are used as herbs.

In addition, the weakness of three hundred seconds (한) and taste is sour (苦辛). It has the effect of fever, mucus, and swelling, so it is difficult to urinate, edema, sediment, seizures, hepatitis, jaundice, mucus, and sociability. Used for symptoms such as injury. The trichophytium contains lignan compounds (sauchinone, saucerneol, manassantin A, manassantin B), flavonoids (rutin, hyperoside, quercitrin and quercetin) and alkaloids (aristolactam BII), which have strong antioxidant effects and apoptosis against cancer cells. apoptosis), etc. (Kim et al., 2011). In particular, Sauserneol D (leaserneol D), a lignan compound, is known to inhibit melanin production, have an antioxidant effect, an anti-asthma effect, and the like.

On the other hand, the present inventors confirmed that the three hundred seconds extract and the compound isolated therefrom inhibits the arginase activity, the three hundred second extract and the compound isolated therefrom increase the NO production, such as atherosclerosis, hypertension, coronary artery disease The present invention has been completed by revealing that it can be used as a therapeutic agent for diseases.

On the other hand, the compounds of 300 seconds derived from arteriosclerosis have a therapeutic effect (3 300 seconds of arteriosclerosis prevention effect and functional food development, Kyungpook National University, 2004) and Korean Patent No. 739398 In the above prior documents, compounds having the same chemical structure as those of the compounds of the present invention are not disclosed.

Korean Registered Patent No. 739398 (Composition for the Prevention and Treatment of Inflammation, Allergic Diseases or Vascular Diseases Containing SC25500 Compound Isolated from the Extract of Triticales, 2007.07.13.

Psychiatry, etc., Prevention of Arteriosclerosis and Development of Functional Foods of Three hundred Seconds, Kyungpook National University, 2004. Xie, Y. W., Ming, D. S., Xu, H. X., Dong, H., But, P. P., Vasorelaxing effects of Caesalpinia sappan involvement of endogenous nitric oxide., Life. Sci., 2000, 67, 1913--1918. Morris, C.R., Poljakovic, M., Lavrisha, L. et al., Decreased arginine bioavailability and increased serum arginase activity in asthma., Am. J. Respir. Crit. Care. Med., 2004, 170, 148-153. Modolell, M., Corraliza, I. M., Link, F., Soler. G., Eichmann, K., Reciprocal regulation of the nitric oxide synthase / arginase balance in mouse bone marrow-derived macrophages by TH1 and TH2 cytokines., Eur. J. Immunol., 1995, 25, 1101-1104. Ryoo, S., Lemmon, C.A., Soucy, K.G. et al., Oxidized low-density lipoprotein-dependent endothelial arginase II activation contributes to impaired nitric oxide signaling., Circ. Res., 2006, 99, 951-960. Nelin, L.D., Wang, X., Zhao, Q. et al., MKP-1 switches arginine metabolism from nitric oxide synthase to arginase following endotoxin challenge., Am. J. Physiol. Cell. Physiol., 2007, 293, 632-640. Faffe, D.S., Flynt, L., Mellema, M. 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. Ryoo, S., Gupta, G., Benjo, A. et al., Endothelial arginase II: a novel target for the treatment of atherosclerosis., Circ. Res., 2008, 102, 923-932. Kim, H.Y. et al., A methylene chloride fraction of Saururus chinensis induces apoptosis through the activation of caspase-3 in prostate and breast cancer cells., Phytomedicine, 2011, 18, 567-574. Woo, A., Min, B., Ryoo, S., 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 is 300 seconds ( Saururus chinensis ) to provide a composition for the treatment or prevention of vascular diseases containing an extract or a compound isolated therefrom.

The present invention provides a method for the prevention of vascular diseases containing Saururus chinensis extract containing at least one compound of saucchinone of Formula 1 and 7-hydroxysauchine of Formula 2, or A therapeutic composition.

[Formula 1]

(2)

The three hundred seconds extract may be prepared by extracting three hundred seconds with a C 1-4 lower alcohol or 10-95% alcohol aqueous solution thereof.

The vascular disease may be a disease selected from the group consisting of arteriosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, heart failure, complications arising after cardiovascular artery arthroplasty, cerebral infarction, cerebral hemorrhage, and stroke.

The present invention also relates to a pharmaceutical composition for the prevention or treatment of vascular diseases containing at least one compound of sacchinone and 7-hydroxysucchin.

The present invention also relates to a dietary supplement for the prevention or improvement of vascular diseases containing at least one compound of the three hundred sec extract, or sacchinone and 7-hydroxysucchin.

Hereinafter, the present invention will be described in more detail.

Preferably, the three hundred seconds extract of the present invention is prepared by distilling under reduced pressure by adding an aqueous solution of 1 to 4 carbon atoms or 1 to 4% lower alcohol having 2 to 20 times (w / v) of the weight of three hundred seconds to three hundred seconds can do. In addition, after adding 10 to 50 times (w / v) of water to the three hundred s. Extract, again 5 to 20 times (w / v) of n-hexane, ethyl acetate, n-butanol of the crude extract weight After purification and fractionation, the compounds that modulate the activity of Arginase II can be separated using chromatography or the like.

The extract of the three hundred seconds, if necessary, extraction with an organic solvent (alcohol, ether, acetone, etc.), distribution of the organic solvent (n-hexane, ethyl acetate, n-butanol) and water, the method by column chromatography, plant components Known methods used for separation extraction and the like can be further purified alone or in combination as appropriate, and can be further purified according to a commercial method as necessary.

The chromatography used in the present invention includes silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, thin layer chromatography Thin layer chromatography (TLC), medium pressure liquid chromatography, and high performance liquid chromatography may be used.

In another aspect, the present invention provides a pharmaceutical composition for the prevention or treatment of vascular diseases containing at least one compound of the three hundred sec extract or Sauchinone and 7-hydroxysachin isolated therefrom.

Pharmaceutical compositions for the prevention and treatment of vascular diseases containing at least one compound of the three hundred sec extract or sachinone and 7-hydroxysucchin according to the present invention, respectively, powders, granules, Oral formulations such as tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions. Carriers, excipients, and diluents that may be included in the composition containing one or more compounds of the three hundred sec extract or sacchinone and 7-hydroxysucchin separated therefrom 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, propylhydroxybenzo 8, 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 preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may be added to at least one compound of sachinone and 7-hydroxysachin, which are separated from the three hundred sec extract. At least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like is mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . 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. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The dosage of the at least one compound of the trichoola extract or the sacchinone and 7-hydroxysucchin isolated therefrom is determined by the age, sex, weight of the subject to be treated and the specific disease or pathological condition, disease or pathology to be treated. It will depend on the severity, route of administration and 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. One or more compounds of the trichophytium extract of the present invention or the sacchinone and 7-hydroxysucchin isolated therefrom can be administered to mammals such as rats, livestock, humans and the like by 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, at least one compound of the three hundred sec extract of the present invention or sachinone and 7-hydroxysucchin isolated therefrom is a composition derived from natural products, so there is almost no toxicity and side effects, so it can be used safely for long-term use for prophylactic purposes. It is a drug that can.

In addition, the present invention provides a health functional food for the prevention or improvement of vascular diseases containing the three hundred seconds extract. Food-acceptable food-aid additives may be added to the health functional food. The health functional food may be in the form of tablets, capsules, pills or liquids. The health functional foods include dairy products such as drinks, meat, sausage, bread, candy, snacks, noodles, It may include nutritional products, including beverages, alcoholic beverages and vitamin complexes, including ionic beverages.

The present invention is directed to a composition for the prevention and treatment of vascular diseases containing at least one compound of Saururus chinensis extract, or Sauchinone and 7-hydroxysauchine isolated therefrom. As related, the trichophytium extract, or one or more compounds of sacchinone and 7-hydroxysucchin are excellent in inhibiting the activity of arginase in vascular cells, such as atherosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic It can be usefully used for the prevention and treatment of vascular diseases such as heart disease, heart failure, complications occurring after carotid angioplasty, cerebral infarction, cerebral hemorrhage, and stroke.

1 is a schematic diagram showing the function of arginase II in vascular cells.

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, it is to be provided so that the contents introduced herein may be thorough and complete and to fully convey the spirit of the invention to those skilled in the art.

< Example  1.Preparation of Extract of Triticale and Sauuccinon  And 7- Of hydroxysachin  Separation>

Three hundred seconds ( Saururus chinensis ) was purchased in May 2010 at Yangnyeong-si, the traditional market in Daegu. Methanol extract (1.5 kg) was obtained by distilling under reduced pressure distillation under reduced pressure three times for 4 hours with 4 L of methanol while reflux-cooled 12 kg of dried three hundred seconds. The methanol extract was suspended in 4 L of water and extracted three times with 4 L of solvent each using n-hexane, ethyl acetate and n-butanol sequentially, and n-hexane fraction (400.8 g), ethyl acetate fraction (584.93 g). ), n-butanol fraction (314.2 g) was obtained. Thereafter, ethyl acetate fraction (584.93 g) was subjected to chromatography on silica gel [CHCl ₃ -MeOH (50: 1 → 5: 1) gradient] to obtain 15 small fractions (Fr.1 to Fr.15). , Fr.7 subfraction having good activity of arginase II was chromatographed using silica gel [hexane-acetone (5: 1 → 0: 1) gradient] to obtain Fr.7-1 (20g) to Fr A small fraction of .7-3 was obtained. Thereafter, the small fraction of Fr.7-1 was chromatographed using silica gel [hexane-EtOAc (10: 1 → 1: 1) gradient] to obtain compound 1 (sauchinone; 120 mg). Fr. The small fraction (25 g) of 7-2 was subjected to chromatography using silica gel [hexane-EtOAc (5: 1 → 1: 1) concentration gradient] to obtain Fr.7-2-1 to Fr.7-2-7. Seven small fractions were obtained, and the small fraction of Fr.7-2-6 was subjected to medium pressure liquid chromatography (MPLC) using an ODS (reversed phase) column [MeOH-H ₂ O (3: 1) Concentration gradient], Compound 2 (7-hydroxysauchinone, 10.8 mg) was isolated.

< Example  2. Sauuccinon  And 7- Of hydroxysachin  Confirmation of Physicochemical Properties>

The properties of the compound isolated in Example 1 were analyzed using ¹ H-NMR and ¹³ C-NMR, and the physicochemical properties of the compound were as follows.

Example  2-1. Compound 1: Succinone ( sauchinone )

White amorphous powder.

85.0 (c 0.1, CHCl₃).-[α]

85.0 ( c 0.1, CHCl ₃ ).

UV (CHCl ₃ ) λ _max nm: 244, 300.

IR ν _max cm ^-1 : 3020, 1214, 748.

EIMS (rel. Int.) M / z : 356 [M] ⁺ (100), 270 (13), 257 (15), 205 (20), 175 (25).

¹ H-NMR (400 MHz, CDCl ₃ ) δ : 0.74 (3H, d, J = 7.6 Hz, H-9 ′), 1.21 (3H, d, J = 7.6 Hz, H-9), 1.67 (1H, m, H-7 ′ eq ), 1.94 (1H, m, H-7 ′ ax ), 1.91 (1H, m, H-8 ′), 2.45 (1H, m, H-8), 2.50 (1H, m , H-1 ′), 2.56 (1H, dd, J = 2.8, 12 Hz, H-6 ′), 3.05 (1H, d, J = 4.8 Hz, H-7), 5.52 (1H, s, H-3 ′), 6.40 (1H, s, H-3), 6.85 (1H, s, H-6), 5.62 and 5.68 (each 1H, s, 4 ′, 5′-OCH ₂ O-), 5.89 and 5.93 ( each 1 H, s, 4,5-OCH ₂ O-).

¹³ C-NMR (100 MHz, CDCl ₃ ) δ : 20.9 (C-9 ′), 21.4 (C-9), 25.3 (C-7 ′), 33.6 (C-8 ′), 34.9 (C-7) , 35.1 (C-8), 35.2 (C-6 ′), 37.6 (C-1 ′), 98.7 (4 ′, 5′-OCH ₂ O-), 99.3 (C-3), 100.3 (C-5 ′), 100.5 (C-3 ′), 101.4 (4,5-OCH ₂ O-), 106.6 (C-6), 115.8 (C-1), 143.3 (C-4), 145.1 (C-2) , 146.8 (C-5), 168.7 (C-4 ′), 199.7 (C-2 ′).

Example  2-2. Compound 2: 7-hydroxysuccin (7- hydroxysauchine )

White amorphous powder.

- 9.46 (c 0.37, CHCl₃).-[α]

9.46 ( c 0.37, CHCl ₃ ).

UV (CHCl ₃ ) λ _max nm: 244, 297.

IR (KBr) ν _max cm ^-1 : 3695, 3019, 1214, 1054, 748.

HR-EIMS (rel. Int.) M / e : 372.1209 [M] ⁺ (100), 373.1247 (22) (calcd for C ₂₀ H ₂₀ O ₇ ; 372.1209).

¹ H-NMR (400 MHz, CDCl ₃ ) δ : 0.60 (3H, d, J = 7.6 Hz, H-9 ′), 1.25 (3H, d, J = 7.2 Hz, H-9), 1.74 (1H, m, H-7 ′ ax ), 1.84 (1H, m, H-7 ′ eq ), 2.06 (1H, m, H-8 ′), 2.50 (1H, m, H-8), 2.55 (1H, m , H-1 ′), 2.60 (1H, d, J = 13.2 Hz, H-6 ′), 5.56 (1H, s, H-3 ′), 6.43 (1H, s, H-3), 7.0 (1H , s, H-6), 5.97 and 5.94 (each 1H, d, J = 1.2HZ, 4 ′, 5′-OCH ₂ O-), 5.68 and 5.67 (each 1H, s, 4,5-OCH ₂ O -).

¹³ C-NMR (100 MHz, CDCl ₃ ) δ : 16.8 (C-9), 20.5 (C-9 ′), 24.4 (C-7 ′), 34.9 (C-8 ′), 40.2 (C-8) , 40.5 (C-1 ′), 44.7 (C-6 ′), 68.9 (C-7), 99.3 (C-3), 100.7 (C-3 ′), 100.9 (C-4 ′), 106.0 (C -6), 119.1 (C-1), 143.8 (C-4), 144.4 (C-2), 148.7 (C-5), 168.2 (C-4 '), 198.6 (C-2'). 101.8 (4,5-OCH ₂ O-), 98.7 (4 ', 5'-OCH ₂ O-).

< Example  3. Confirmation of Arginase Inhibitory Effect>

Arginase activity assays are described in Ryoo et. al ., 2006]. Arginase is in the isomeric form of arginase I and II (Ryoo et. al ., 2010), with reference to this, it was confirmed the activity of arginase II in the cell eluate of human umbilical vein endothelial cells (HUVEC). As a control of this experiment, lysates of kidney cells of mice treated with DMSO, which is a solvent of a sample (three hundred seconds methanol extract, sautinone, and 7-hydroxysuccin) in the same volume as the sample usage were used.

To this end, HUVEC was treated with a cell lysis buffer (50 mM Tris-HCl, pH 7.5, 0.1 mM EDTA and protease inhibitors) and homogenized at 4 ° C. Centrifugation was carried out at 14,000 x g and the supernatant was taken to obtain the HUVEC eluate. It was. The HUVEC eluate was diluted 10-fold with 50 mM tris-HCl, and 33.5 μl of 50 mM Tris-HC, sample (three hundred seconds methanol extract, sautinone and 7-hydroxysauchin: 1 mg / μl of the sample 2, 4 , 8, 16, 32, 50 μl each), 10 μl diluted HUVEC eluate, 25 μl L-arginine (L-arginine, 0.1M) was added. Thereafter, the reaction was carried out at 37 ° C. for 1 hour, and then the reaction was stopped with an acid solution (H ₂ SO ₄ : H ₃ PO ₄ : DW = 1: 3: 7).

Next, 25 μl of INSP (9% α-isonitrosopropiophenone in absolute ethanol) was added and color developed at 97 ° C. for 45 minutes (color reaction with urea). After 10 minutes of dark reaction, absorbance was measured at 550 nm. In addition, the absorbance values were compared with the control group was not treated. Enzyme activity (pmol Urea / min / mg protein) was calculated from the urea standard curve, and enzyme activity inhibition was measured as a percentage of the enzyme activity in the absence of the sample by measuring the arginase activity with the sample. The concentration of each sample showing 50% activity inhibition (the half maximal inhibitory concentration (IC ₅₀ )) is shown in Table 1 below.

Referring to Table 1, it can be seen that the three hundred seconds methanol extract, Sauchinone and 7-hydroxysucchin prepared or separated in Example 1 has the effect of inhibiting the activity of arginase II.

Condition IC ₅₀ in HUVEC Eluates (μM) Sauuccinon 61.4 7-hydroxysachin 89.6 300 seconds methanol extract 300> Positive Control: PG 1.0 ※ PG: Piceatannol-3'- O­β -D-glucopyranosi

< Example  4. Toxicity Test>

Example  4-1. Acute toxicity

This study was conducted to investigate the toxicity of acutely (within 24 hours) to an animal body and to determine the mortality rate when a short-term overdose of methanol extract, sautinone and 7-hydroxysucchin of the present invention was ingested. Was performed. 40 ICR mouse strains, which are common mice, were assigned to 10 control groups and 10 experimental groups. Nothing was administered to the control group, and the experimental group was 2.0 g / kg (300 times the amount used in the general animal experiments) of the 300 seconds methanol extract prepared in Example 1, and sacchinone and 7-hydroxysaucine. It was administered orally at the concentration of. The mortality was examined 24 hours after administration, and the control group and the experimental group treated with 300 g of methanol extract, sacchinone, and 7-hydroxysachin at 2.0 g / kg concentration survived.

Example  4-2. Experimental group  And control organ organs and tissue toxicity experiments

In order to investigate the effects on the organs (tissues) of animals in C57BL / 6J mice, only the experimental group and the solvent administered with the 300 sec methanol extract, sautinone, and 7-hydroxysachin prepared and separated in Example 1 Blood was collected after 8 weeks from animals in the control group, and blood concentrations of glutamate-pyruvate transferase (GPT) and blood urea nitrogen (BUN) were measured using a Select E (Vital Scientific NV, Netherland) instrument. 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.

< Examples  1. Pharmaceutical Formulation example >

Examples  1-1. Manufacture of tablets

20 g of sacchinone isolated in Example 1 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.

Examples  1-2. Injection preparation

2 g of sacchinone separated in Example 1 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.

< Examples  2. Food Manufacturing example >

Examples  2-1. Manufacture of cooking seasonings

Health care cooking seasoning was prepared by using 0.2 to 10% by weight of the methanol extract of 300 seconds prepared in Example 1.

Examples  2-2. Manufacture of flour food products

The triglyceride methanol extract prepared in Example 1 was added to the flour at 0.1 to 5.0% by weight, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare foods for health promotion.

Examples  2-3. soup  And juicy ( gravies )

The 300 ml methanol extract prepared in Example 1 was added in an amount of 0.1 to 1.0% by weight to soups and broths to prepare meat products for health promotion, soups and noodles for noodles.

Examples  2-4. dairy product( dairy products )

The methanol extract of 300 seconds prepared in Example 1 was added to milk at 0.1 to 1.0% by weight, and various dairy products such as butter and ice cream were prepared using the milk.

< Examples  3. Drink Manufacturing example >

Examples  3-1. Vegetable juice  Produce

0.5 g of the triticale methanol extract prepared in Example 1 was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

Examples  3-2. Fruit juice  Produce

0.1 g of the triticale methanol extract prepared in Example 1 was added to 1,000 ml of apple or grape juice to prepare fruit juice for health promotion.

Claims (3)

Arteriosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, heart failure, cerebral infarction, cerebral hemorrhage, and stroke containing 7-hydroxysauchine of formula (2) Prophylactic or therapeutic pharmaceutical composition.
(2)

delete 7-hydroxysauchine compound of the formula (2).
(2)

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